EP3693025A1 - Adenoassoziierte virus-virionen mit veränderlichem kapsid und verwendungsverfahren dafür - Google Patents

Adenoassoziierte virus-virionen mit veränderlichem kapsid und verwendungsverfahren dafür Download PDF

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Publication number
EP3693025A1
EP3693025A1 EP20151479.1A EP20151479A EP3693025A1 EP 3693025 A1 EP3693025 A1 EP 3693025A1 EP 20151479 A EP20151479 A EP 20151479A EP 3693025 A1 EP3693025 A1 EP 3693025A1
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Prior art keywords
amino acids
aav
seq
cell
capsid protein
Prior art date
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EP20151479.1A
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English (en)
French (fr)
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EP3693025B1 (de
Inventor
David V. Schaffer
Ryan R. Klimczak
James T. KOERBER
John G. Flannery
Deniz DALKARA MOUROT
Meike VISEL
Leah C.T. BYRNE
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University of California
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University of California
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Priority to PL20151479T priority Critical patent/PL3693025T3/pl
Priority to HRP20220036TT priority patent/HRP20220036T1/hr
Priority to EP21201945.9A priority patent/EP4005603A1/de
Priority to SI201231974T priority patent/SI3693025T1/sl
Priority to RS20220014A priority patent/RS62795B1/sr
Publication of EP3693025A1 publication Critical patent/EP3693025A1/de
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Publication of EP3693025B1 publication Critical patent/EP3693025B1/de
Priority to CY20221100029T priority patent/CY1124905T1/el
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    • C12N2750/14151—Methods of production or purification of viral material
    • C12N2750/14152—Methods of production or purification of viral material relating to complementing cells and packaging systems for producing virus or viral particles
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2810/00—Vectors comprising a targeting moiety
    • C12N2810/40—Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/16—Primer sets for multiplex assays

Definitions

  • Photoreceptors are the first neurons in the retina to receive and process visual information, converting visible electromagnetic radiation into hyperpolarized responses through phototransduction.
  • the overwhelming majority of inherited retinal diseases result in the loss of these cells, either directly, such as in dominant mutations that affect rhodopsin protein folding, or indirectly, such as in recessive mutations that affect retinal recycling pathways in the retinal pigment epithelium (RPE).
  • RPE retinal pigment epithelium
  • AAV belongs to the Parvoviridae family and Dependovirus genus, whose members require co-infection with a helper virus such as adenovirus to promote replication, and AAV establishes a latent infection in the absence of a helper.
  • Virions are composed of a 25 nm icosahedral capsid encompassing a 4.9 kb single-stranded DNA genome with two open reading frames: rep and cap.
  • the non-structural rep gene encodes four regulatory proteins essential for viral replication, whereas cap encodes three structural proteins (VP1-3) that assemble into a 60-mer capsid shell.
  • This viral capsid mediates the ability of AAV vectors to overcome many of the biological barriers of viral transduction-including cell surface receptor binding, endocytosis, intracellular trafficking, and unpackaging in the nucleus.
  • the present disclosure provides adeno-associated virus (AAV) virions with altered capsid protein, where the AAV virions exhibit greater infectivity of a retinal cell, when administered via intravitreal injection, compared to wild-type AAV.
  • AAV adeno-associated virus
  • the present disclosure further provides methods of delivering a gene product to a retinal cell in an individual, and methods of treating ocular disease.
  • retinal cell can refer herein to any of the cell types that comprise the retina, such as retinal ganglion cells, amacrine cells, horizontal cells, bipolar cells, and photoreceptor cells including rods and cones, Müller glial cells, and retinal pigmented epithelium.
  • AAV is an abbreviation for adeno-associated virus, and may be used to refer to the virus itself or derivatives thereof. The term covers all subtypes and both naturally occurring and recombinant forms, except where required otherwise.
  • the abbreviation “rAAV” refers to recombinant adeno-associated virus, also referred to as a recombinant AAV vector (or "rAAV vector”).
  • AAV includes AAV type 1 (AAV-1), AAV type 2 (AAV-2), AAV type 3 (AAV-3), AAV type 4 (AAV-4), AAV type 5 (AAV-5), AAV type 6 (AAV-6), AAV type 7 (AAV-7), AAV type 8 (AAV-8), avian AAV, bovine AAV, canine AAV, equine AAV, primate AAV, non-primate AAV, and ovine AAV.
  • Primary AAV refers to AAV that infect primates
  • non-primate AAV refers to AAV that infect non-primate mammals
  • bovine AAV refers to AAV that infect bovine mammals, etc.
  • rAAV vector refers to an AAV vector comprising a polynucleotide sequence not of AAV origin (i.e., a polynucleotide heterologous to AAV), typically a sequence of interest for the genetic transformation of a cell.
  • the heterologous polynucleotide is flanked by at least one, and generally by two, AAV inverted terminal repeat sequences (ITRs).
  • ITRs AAV inverted terminal repeat sequences
  • the term rAAV vector encompasses both rAAV vector particles and rAAV vector plasmids.
  • An rAAV vector may either be single-stranded (ssAAV) or self-complementary (scAAV).
  • An "AAV virus” or “AAV viral particle” or “rAAV vector particle” refers to a viral particle composed of at least one AAV capsid protein (typically by all of the capsid proteins of a wild-type AAV) and an encapsidated polynucleotide rAAV vector. If the particle comprises a heterologous polynucleotide (i.e. a polynucleotide other than a wild-type AAV genome such as a transgene to be delivered to a mammalian cell), it is typically referred to as an "rAAV vector particle” or simply an "rAAV vector". Thus, production of rAAV particle necessarily includes production of rAAV vector, as such a vector is contained within an rAAV particle.
  • Packaging refers to a series of intracellular events that result in the assembly and encapsidation of an AAV particle.
  • AAV "rep” and “cap” genes refer to polynucleotide sequences encoding replication and encapsidation proteins of adeno-associated virus. AAV rep and cap are referred to herein as AAV "packaging genes.”
  • helper virus for AAV refers to a virus that allows AAV (e.g. wild-type AAV) to be replicated and packaged by a mammalian cell.
  • helper viruses for AAV are known in the art, including adenoviruses, herpesviruses and poxviruses such as vaccinia.
  • the adenoviruses encompass a number of different subgroups, although Adenovirus type 5 of subgroup C is most commonly used.
  • Numerous adenoviruses of human, non-human mammalian and avian origin are known and available from depositories such as the ATCC.
  • Viruses of the herpes family include, for example, herpes simplex viruses (HSV) and Epstein-Barr viruses (EBV), as well as cytomegaloviruses (CMV) and pseudorabies viruses (PRV); which are also available from depositories such as ATCC.
  • HSV herpes simplex viruses
  • EBV Epstein-Barr viruses
  • CMV cytomegaloviruses
  • PRV pseudorabies viruses
  • Helper virus function(s) refers to function(s) encoded in a helper virus genome which allow AAV replication and packaging (in conjunction with other requirements for replication and packaging described herein).
  • helper virus function may be provided in a number of ways, including by providing helper virus or providing, for example, polynucleotide sequences encoding the requisite function(s) to a producer cell in trans.
  • a plasmid or other expression vector comprising nucleotide sequences encoding one or more adenoviral proteins is transfected into a producer cell along with an rAAV vector.
  • infectious virus or viral particle is one that comprises a competently assembled viral capsid and is capable of delivering a polynucleotide component into a cell for which the viral species is tropic.
  • the term does not necessarily imply any replication capacity of the virus.
  • Assays for counting infectious viral particles are described elsewhere in this disclosure and in the art.
  • Viral infectivity can be expressed as the ratio of infectious viral particles to total viral particles. Methods of determining the ratio of infectious viral particle to total viral particle are known in the art. See, e.g., Grainger et al. (2005) Mol. Ther. 11:S337 (describing a TCID50 infectious titer assay); and Zolotukhin et al. (1999) Gene Ther. 6:973 . See also the Examples.
  • a “replication-competent” virus refers to a phenotypically wild-type virus that is infectious, and is also capable of being replicated in an infected cell (i.e. in the presence of a helper virus or helper virus functions).
  • replication competence generally requires the presence of functional AAV packaging genes.
  • rAAV vectors as described herein are replication-incompetent in mammalian cells (especially in human cells) by virtue of the lack of one or more AAV packaging genes.
  • rAAV vector preparations as described herein are those which contain few if any replication competent AAV (rcAAV, also referred to as RCA) (e.g., less than about 1 rcAAV per 10 2 rAAV particles, less than about 1 rcAAV per 10 4 rAAV particles, less than about 1 rcAAV per 10 8 rAAV particles, less than about 1 rcAAV per 10 12 rAAV particles, or no rcAAV).
  • rcAAV also referred to as RCA
  • polynucleotide refers to a polymeric form of nucleotides of any length, including deoxyribonucleotides or ribonucleotides, or analogs thereof.
  • a polynucleotide may comprise modified nucleotides, such as methylated nucleotides and nucleotide analogs, and may be interrupted by non-nucleotide components. If present, modifications to the nucleotide structure may be imparted before or after assembly of the polymer.
  • polynucleotide refers interchangeably to double- and single-stranded molecules. Unless otherwise specified or required, any embodiment of the invention described herein that is a polynucleotide encompasses both the double-stranded form and each of two complementary single-stranded forms known or predicted to make up the double-stranded form.
  • Nucleic acid hybridization reactions can be performed under conditions of different "stringency”. Conditions that increase stringency of a hybridization reaction of widely known and published in the art. See, e.g., Sambrook et al. Molecular Cloning, A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989 , herein incorporated by reference. For example, see page 7.52 of Sambrook et al.
  • Examples of relevant conditions include (in order of increasing stringency): incubation temperatures of 25°C, 37°C, 50°C and 68°C; buffer concentrations of 10 x SSC, 6 x SSC, 1 x SSC, 0.1 x SSC (where 1 x SSC is 0.15 M NaCl and 15 mM citrate buffer) and their equivalents using other buffer systems; formamide concentrations of 0%, 25%, 50%, and 75%; incubation times from 5 minutes to 24 hours; 1, 2, or more washing steps; wash incubation times of 1, 2, or 15 minutes; and wash solutions of 6 x SSC, 1 x SSC, 0.1 x SSC, or deionized water.
  • stringent hybridization conditions hybridization at 50°C or higher and 0.1 ⁇ SSC (15 mM sodium chloride/1.5 mM sodium citrate). Another example of stringent hybridization conditions is overnight incubation at 42°C in a solution: 50% formamide, 1 ⁇ SSC (150 mM NaCl, 15 mM sodium citrate), 50 mM sodium phosphate (pH 7.6), 5 ⁇ Denhardt's solution, 10% dextran sulfate, and 20 ⁇ g/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1 ⁇ SSC at about 65°C.
  • stringent hybridization conditions comprise: prehybridization for 8 hours to overnight at 65° C in a solution comprising 6X single strength citrate (SSC) (1X SSC is 0.15 M NaCl, 0.015 M Na citrate; pH 7.0), 5X Denhardt's solution, 0.05% sodium pyrophosphate and 100 ⁇ g/ml herring sperm DNA; hybridization for 18-20 hours at 65° C in a solution containing 6X SSC, 1X Denhardt's solution, 100 ⁇ g/ml yeast tRNA and 0.05% sodium pyrophosphate; and washing of filters at 65° C for 1 h in a solution containing 0.2X SSC and 0.1% SDS (sodium dodecyl sulfate).
  • SSC single strength citrate
  • Stringent hybridization conditions are hybridization conditions that are at least as stringent as the above representative conditions. Other stringent hybridization conditions are known in the art and may also be employed to identify nucleic acids of this particular embodiment of the invention.
  • T m is the temperature in degrees Celsius at which 50% of a polynucleotide duplex made of complementary strands hydrogen bonded in anti-parallel direction by Watson-Crick base pairing dissociates into single strands under conditions of the experiment.
  • [X + ] is the cation concentration (usually sodium ion, Na + ) in mol/L
  • (%G/C) is the number of G and C residues as a percentage of total residues in the duplex
  • (%F) is the percent formamide in solution (wt/vol); and L is the number of nucleotides in each strand of the duplex.
  • a polynucleotide or polypeptide has a certain percent "sequence identity" to another polynucleotide or polypeptide, meaning that, when aligned, that percentage of bases or amino acids are the same when comparing the two sequences. Sequence similarity can be determined in a number of different manners. To determine sequence identity, sequences can be aligned using the methods and computer programs, including BLAST, available over the world wide web at ncbi.nlm.nih.gov/BLAST/. Another alignment algorithm is FASTA, available in the Genetics Computing Group (GCG) package, from Madison, Wisconsin, USA, a wholly owned subsidiary of Oxford Molecular Group, Inc. Other techniques for alignment are described in Methods in Enzymology, vol.
  • GCG Genetics Computing Group
  • the program has default parameters determined by the sequences inputted to be compared.
  • the sequence identity is determined using the default parameters determined by the program. This program is available also from Genetics Computing Group (GCG) package, from Madison, Wisconsin, USA.
  • GCG Genetics Computing Group
  • FastDB is described in Current Methods in Sequence Comparison and Analysis, Macromolecule Sequencing and Synthesis, Selected Methods and Applications, pp. 127-149, 1988, Alan R. Liss, Inc. Percent sequence identity is calculated by FastDB based upon the following parameters: Mismatch Penalty: 1.00; Gap Penalty: 1.00; Gap Size Penalty: 0.33; and Joining Penalty: 30.0.
  • a “gene” refers to a polynucleotide containing at least one open reading frame that is capable of encoding a particular protein after being transcribed and translated.
  • a “gene product” is a molecule resulting from expression of a particular gene.
  • Gene products include, e.g., a polypeptide, an aptamer, an interfering RNA, an mRNA, and the like.
  • a “small interfering” or “short interfering RNA” or siRNA is a RNA duplex of nucleotides that is targeted to a gene interest (a "target gene”).
  • An “RNA duplex” refers to the structure formed by the complementary pairing between two regions of a RNA molecule.
  • siRNA is "targeted” to a gene in that the nucleotide sequence of the duplex portion of the siRNA is complementary to a nucleotide sequence of the targeted gene.
  • the length of the duplex of siRNAs is less than 30 nucleotides.
  • the duplex can be 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11 or 10 nucleotides in length.
  • the length of the duplex is 19-25 nucleotides in length.
  • the RNA duplex portion of the siRNA can be part of a hairpin structure.
  • the hairpin structure may contain a loop portion positioned between the two sequences that form the duplex.
  • the loop can vary in length. In some embodiments the loop is 5, 6, 7, 8, 9, 10, 11, 12 or 13 nucleotides in length.
  • the hairpin structure can also contain 3' or 5' overhang portions. In some embodiments, the overhang is a 3' or a 5' overhang 0, 1, 2, 3, 4 or 5 nucleotides in length.
  • shRNA is a polynucleotide construct that can be made to express an interfering RNA such as siRNA.
  • Recombinant as applied to a polynucleotide means that the polynucleotide is the product of various combinations of cloning, restriction or ligation steps, and other procedures that result in a construct that is distinct from a polynucleotide found in nature.
  • a recombinant virus is a viral particle comprising a recombinant polynucleotide. The terms respectively include replicates of the original polynucleotide construct and progeny of the original virus construct.
  • control element or "control sequence” is a nucleotide sequence involved in an interaction of molecules that contributes to the functional regulation of a polynucleotide, including replication, duplication, transcription, splicing, translation, or degradation of the polynucleotide. The regulation may affect the frequency, speed, or specificity of the process, and may be enhancing or inhibitory in nature.
  • Control elements known in the art include, for example, transcriptional regulatory sequences such as promoters and enhancers.
  • a promoter is a DNA region capable under certain conditions of binding RNA polymerase and initiating transcription of a coding region usually located downstream (in the 3' direction) from the promoter.
  • “Operatively linked” or “operably linked” refers to a juxtaposition of genetic elements, wherein the elements are in a relationship permitting them to operate in the expected manner. For instance, a promoter is operatively linked to a coding region if the promoter helps initiate transcription of the coding sequence. There may be intervening residues between the promoter and coding region so long as this functional relationship is maintained.
  • An "expression vector” is a vector comprising a region which encodes a polypeptide of interest, and is used for effecting the expression of the protein in an intended target cell.
  • An expression vector also comprises control elements operatively linked to the encoding region to facilitate expression of the protein in the target.
  • the combination of control elements and a gene or genes to which they are operably linked for expression is sometimes referred to as an "expression cassette,” a large number of which are known and available in the art or can be readily constructed from components that are available in the art.
  • Heterologous means derived from a genotypically distinct entity from that of the rest of the entity to which it is being compared.
  • a polynucleotide introduced by genetic engineering techniques into a plasmid or vector derived from a different species is a heterologous polynucleotide.
  • a promoter removed from its native coding sequence and operatively linked to a coding sequence with which it is not naturally found linked is a heterologous promoter.
  • an rAAV that includes a heterologous nucleic acid encoding a heterologous gene product is an rAAV that includes a nucleic acid not normally included in a naturally-occurring, wild-type AAV, and the encoded heterologous gene product is a gene product not normally encoded by a naturally-occurring, wild-type AAV.
  • genetic alteration and “genetic modification” (and grammatical variants thereof), are used interchangeably herein to refer to a process wherein a genetic element (e.g., a polynucleotide) is introduced into a cell other than by mitosis or meiosis.
  • the element may be heterologous to the cell, or it may be an additional copy or improved version of an element already present in the cell.
  • Genetic alteration may be effected, for example, by transfecting a cell with a recombinant plasmid or other polynucleotide through any process known in the art, such as electroporation, calcium phosphate precipitation, or contacting with a polynucleotide-liposome complex.
  • Genetic alteration may also be effected, for example, by transduction or infection with a DNA or RNA virus or viral vector.
  • the genetic element is introduced into a chromosome or mini-chromosome in the cell; but any alteration that changes the phenotype and/or genotype of the cell and its progeny is included in this term.
  • a cell is said to be “stably” altered, transduced, genetically modified, or transformed with a genetic sequence if the sequence is available to perform its function during extended culture of the cell in vitro.
  • a cell is "heritably” altered (genetically modified) in that a genetic alteration is introduced which is also inheritable by progeny of the altered cell.
  • polypeptide refers to polymers of amino acids of any length.
  • the terms also encompass an amino acid polymer that has been modified; for example, disulfide bond formation, glycosylation, lipidation, phosphorylation, or conjugation with a labeling component.
  • Polypeptides such as anti-angiogenic polypeptides, neuroprotective polypeptides, and the like, when discussed in the context of delivering a gene product to a mammalian subject, and compositions therefor, refer to the respective intact polypeptide, or any fragment or genetically engineered derivative thereof, which retains the desired biochemical function of the intact protein.
  • references to nucleic acids encoding anti-angiogenic polypeptides, nucleic acids encoding neuroprotective polypeptides, and other such nucleic acids for use in delivery of a gene product to a mammalian subject include polynucleotides encoding the intact polypeptide or any fragment or genetically engineered derivative possessing the desired biochemical function.
  • an "isolated" plasmid, nucleic acid, vector, virus, virion, host cell, or other substance refers to a preparation of the substance devoid of at least some of the other components that may also be present where the substance or a similar substance naturally occurs or is initially prepared from.
  • an isolated substance may be prepared by using a purification technique to enrich it from a source mixture. Enrichment can be measured on an absolute basis, such as weight per volume of solution, or it can be measured in relation to a second, potentially interfering substance present in the source mixture. Increasing enrichments of the embodiments of this disclosure are increasingly more isolated.
  • An isolated plasmid, nucleic acid, vector, virus, host cell, or other substance is in some embodiments purified, e.g., from about 80% to about 90% pure, at least about 90% pure, at least about 95% pure, at least about 98% pure, or at least about 99%, or more, pure.
  • treatment refers to obtaining a desired pharmacologic and/or physiologic effect.
  • the effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for a disease and/or adverse effect attributable to the disease.
  • Treatment covers any treatment of a disease in a mammal, particularly in a human, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease or at risk of acquiring the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease.
  • the terms “individual,” “host,” “subject,” and “patient” are used interchangeably herein, and refer to a mammal, including, but not limited to, human and non-human primates, including simians and humans; mammalian sport animals (e.g., horses); mammalian farm animals (e.g., sheep, goats, etc.); mammalian pets (dogs, cats, etc.); and rodents (e.g., mice, rats, etc.).
  • mammalian sport animals e.g., horses
  • mammalian farm animals e.g., sheep, goats, etc.
  • mammalian pets dogs, cats, etc.
  • rodents e.g., mice, rats, etc.
  • the present disclosure provides adeno-associated virus (AAV) virions with altered capsid protein, where the AAV virions exhibit greater infectivity of a retinal cell, when administered via intravitreal injection, compared to wild-type AAV when administered via intravitreal injection.
  • AAV adeno-associated virus
  • the present disclosure further provides methods of delivering a gene product to a retinal cell in an individual, and methods of treating ocular disease.
  • the retinal cell can be a photoreceptor (e.g., rods; cones), a retinal ganglion cell (RGC), a Müller cell (a Müller glial cell), a bipolar cell, an amacrine cell, a horizontal cell, or a retinal pigmented epithelium (RPE) cell.
  • a photoreceptor e.g., rods; cones
  • RRC retinal ganglion cell
  • Müller cell a Müller glial cell
  • bipolar cell e.g., an amacrine cell
  • amacrine cell e.g., a amacrine cell
  • horizontal cell e.g., a horizontal cell
  • RPE retinal pigmented epithelium
  • the present disclosure provides a variant AAV capsid protein, where the variant AAV capsid protein comprises an insertion of from about 5 amino acids to about 11 amino acids in an insertion site in the capsid protein GH loop or loop IV, relative to a corresponding parental AAV capsid protein, and where the variant capsid protein, when present in an AAV virion, confers increased infectivity of a retinal cell compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • the retinal cell is a photoreceptor cell (e.g., rods; cones).
  • the retinal cell is an RGC.
  • the retinal cell is an RPE cell.
  • the retinal cell is a Müller cell.
  • Other retinal cells include amacrine cells, bipolar cells, and horizontal cells.
  • An "insertion of from about 5 amino acids to about 11 amino acids” is also referred to herein as a "peptide insertion” (e.g., a heterologous peptide insertion).
  • a "corresponding parental AAV capsid protein” refers to an AAV capsid protein of the same AAV serotype, without the peptide insertion.
  • the insertion site is in the GH loop, or loop IV, of the AAV capsid protein, e.g., in a solvent-accessible portion of the GH loop, or loop IV, of the AAV capsid protein.
  • the insertion site can be within amino acids 411-650 of an AAV capsid protein, as depicted in Figures 17A and 17B .
  • the insertion site can be within amino acids 570-611 of AAV2, within amino acids 571-612 of AAV1, within amino acids 560-601 of AAV5, within amino acids 571 to 612 of AAV6, within amino acids 572 to 613 of AAV7, within amino acids 573 to 614 of AAV8, within amino acids 571 to 612 of AAV9, or within amino acids 573 to 614 of AAV10, as depicted in Figure 6 .
  • the insertion site can be between amino acids 587 and 588 of AAV2, or the corresponding positions of the capsid subunit of another AAV serotype. It should be noted that the insertion site 587/588 is based on an AAV2 capsid protein. From about 5 amino acids to about 11 amino acids can be inserted in a corresponding site in an AAV serotype other than AAV2 (e.g., AAV8, AAV9, etc.).
  • the insertion site is a single insertion site between two adjacent amino acids located between amino acids 570-614 of VP1 of any AAV serotype, e.g., the insertion site is between two adjacent amino acids located in amino acids 570-610, amino acids 580-600, amino acids 570-575, amino acids 575-580, amino acids 580-585, amino acids 585-590, amino acids 590-600, or amino acids 600-614, of VP1 of any AAV serotype or variant.
  • the insertion site can be between amino acids 580 and 581, amino acids 581 and 582, amino acids 583 and 584, amino acids 584 and 585, amino acids 585 and 586, amino acids 586 and 587, amino acids 587 and 588, amino acids 588 and 589, or amino acids 589 and 590.
  • the insertion site can be between amino acids 575 and 576, amino acids 576 and 577, amino acids 577 and 578, amino acids 578 and 579, or amino acids 579 and 580.
  • the insertion site can be between amino acids 590 and 591, amino acids 591 and 592, amino acids 592 and 593, amino acids 593 and 594, amino acids 594 and 595, amino acids 595 and 596, amino acids 596 and 597, amino acids 597 and 598, amino acids 598 and 599, or amino acids 599 and 600.
  • the insertion site can be between amino acids 587 and 588 of AAV2, between amino acids 590 and 591 of AAV1, between amino acids 575 and 576 of AAV5, between amino acids 590 and 591 of AAV6, between amino acids 589 and 590 of AAV7, between amino acids 590 and 591 of AAV8, between amino acids 588 and 589 of AAV9, or between amino acids 588 and 589 of AAV10.
  • the insertion site can be between amino acids 450 and 460 of an AAV capsid protein, as shown in Figure 17A .
  • the insertion site can be at (e.g., immediately N-terminal to) amino acid 453 of AAV2, at amino acid 454 of AAV1, at amino acid 454 of AAV6, at amino acid 456 of AAV7, at amino acid 456 of AAV8, at amino acid 454 of AAV9, or at amino acid 456 of AAV10, as shown in Figure 17A .
  • a subject capsid protein includes a GH loop comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence set forth in Figure 18A-C .
  • a peptide of from about 5 amino acids to about 11 amino acids in length is inserted into the GH loop of an AAV capsid.
  • the insertion peptide has a length of 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, or 11 amino acids.
  • the insertion peptide can comprise an amino acid sequence of any one of the formulas set forth below.
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula I: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IIa: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IIb: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula III: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IV: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • the insertion peptide can comprise an amino acid sequence selected from LGETTRP (SEQ ID NO: 13), NETITRP (SEQ ID NO:14), KAGQANN (SEQ ID NO:15), KDPKTTN (SEQ ID NO:16), KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60).
  • LGETTRP SEQ ID NO: 13
  • NETITRP SEQ ID NO:14
  • KAGQANN SEQ ID NO:15
  • KDPKTTN SEQ ID NO:16
  • KDTDTTR SEQ ID NO:57
  • RAGGSVG SEQ ID NO:58
  • AVDTTKF SEQ ID NO:59
  • STGKVPN STGKVPN
  • the insertion peptide has from 1 to 4 spacer amino acids (Y 1 -Y 4 ) at the amino terminus and/or at the carboxyl terminus of any one of LGETTRP (SEQ ID NO: 13), NETITRP (SEQ ID NO:14), KAGQANN (SEQ ID NO:15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60).
  • Suitable spacer amino acids include, but are not limited to, leucine, alanine, glycine, and serine.
  • an insertion peptide has one of the following amino acid sequences: LALGETTRPA (SEQ ID NO:45); LANETITRPA (SEQ ID NO:46), LAKAGQANNA (SEQ ID NO:47), LAKDPKTTNA (SEQ ID NO:48), LAKDTDTTRA (SEQ ID NO:61), LARAGGSVGA (SEQ ID NO:62), LAAVDTTKFA (SEQ ID NO:63), and LASTGKVPNA (SEQ ID NO:64).
  • an insertion peptide has one of the following amino acid sequences: AALGETTRPA (SEQ ID NO:49); AANETITRPA (SEQ ID NO:50), AAKAGQANNA (SEQ ID NO:51), and AAKDPKTTNA (SEQ ID NO:52).
  • an insertion peptide has one of the following amino acid sequences: GLGETTRPA (SEQ ID NO:53); GNETITRPA (SEQ ID NO:54), GKAGQANNA (SEQ ID NO:55), and GKDPKTTNA (SEQ ID NO:56).
  • an insertion peptide comprises one of KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60), flanked on the C-terminus by AA and on the N-terminus by A; or comprises one of KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60) flanked on the C-terminus by G and on the N-terminus by A.
  • a subject variant AAV capsid does not include any other amino acid substitutions, insertions, or deletions, other than an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject variant AAV capsid includes from 1 to about 25 amino acid insertions, deletions, or substitutions, compared to the parental AAV capsid protein, in addition to an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject variant AAV capsid includes from 1 to about 5, from about 5 to about 10, from about 10 to about 15, from about 15 to about 20, or from about 20 to about 25 amino acid insertions, deletions, or substitutions, compared to the parental AAV capsid protein, in addition to an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject variant capsid polypeptide does not include one, two, three, or four, of the following amino acid substitutions: Y273F, Y444F, Y500F, and Y730F.
  • a subject variant capsid polypeptide comprises, in addition to an insertion peptide as described above, one, two, three, or four, of the following amino acid substitutions: Y273F, Y444F, Y500F, and Y730F.
  • a variant AAV capsid polypeptide is a chimeric capsid, e.g., the capsid comprises a portion of an AAV capsid of a first AAV serotype and a portion of an AAV capsid of a second serotype; and comprises an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject variant capsid protein comprises an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99%, amino acid sequence identity to the amino acid sequence provided in Figure 5 ; and an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject variant capsid protein is isolated, e.g., purified.
  • a subject variant capsid protein is included in an AAV vector, which is also provided.
  • a subject variant capsid protein can be included in a recombinant AAV virion.
  • the present disclosure provides a recombinant adeno-associated virus (rAAV) virion comprising: a) a variant AAV capsid protein, where the variant AAV capsid protein comprises an insertion of from about 5 amino acids to about 11 amino acids in an insertion site in the capsid protein GH loop or loop IV, relative to a corresponding parental AAV capsid protein, and where the variant capsid protein confers increased infectivity of a retinal cell compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein; and b) a heterologous nucleic acid comprising a nucleotide sequence encoding a gene product.
  • rAAV adeno-associated virus
  • the retinal cell is a photoreceptor cell (e.g., rods and/or cones). In other cases, the retinal cell is an RGC cell. In other cases, the retinal cell is an RPE cell. In other cases, the retinal cell is a Müller cell. In other cases, retinal cells may include amacrine cells, bipolar cells, and horizontal cells.
  • An "insertion of from about 5 amino acids to about 11 amino acids” is also referred to herein as a "peptide insertion" (e.g., a heterologous peptide insertion).
  • a "corresponding parental AAV capsid protein” refers to an AAV capsid protein of the same AAV serotype, without the peptide insertion.
  • the insertion site is in the GH loop, or loop IV, of the AAV capsid protein, e.g., in a solvent-accessible portion of the GH loop, or loop IV, of the AAV capsid protein.
  • GH loop see, e.g., van Vliet et al. (2006) Mol. Ther. 14:809 ; Padron et al. (2005) J. Virol. 79:5047 ; and Shen et al. (2007) Mol. Ther. 15:1955 .
  • the insertion site is within amino acids 570-611 of AAV2, within amino acids 571-612 of AAV1, within amino acids 560-601 of AAV5, within amino acids 571 to 612 of AAV6, within amino acids 572 to 613 of AAV7, within amino acids 573 to 614 of AAV8, within amino acids 571 to 612 of AAV9, or within amino acids 573 to 614 of AAV10.
  • From about 5 amino acids to about 11 amino acids are inserted in an insertion site in the GH loop or loop IV of the capsid protein relative to a corresponding parental AAV capsid protein.
  • the insertion site can be between amino acids 587 and 588 of AAV2, or the corresponding positions of the capsid subunit of another AAV serotype. It should be noted that the insertion site 587/588 is based on an AAV2 capsid protein. From about 5 amino acids to about 11 amino acids can be inserted in a corresponding site in an AAV serotype other than AAV2 (e.g., AAV8, AAV9, etc.).
  • the insertion site is a single insertion site between two adjacent amino acids located between amino acids 570-614 of VP1 of any AAV serotype, e.g., the insertion site is between two adjacent amino acids located in amino acids 570-614, amino acids 580-600, amino acids 570-575, amino acids 575-580, amino acids 580-585, amino acids 585-590, amino acids 590-600, or amino acids 600-610, of VP1 of any AAV serotype or variant.
  • the insertion site can be between amino acids 580 and 581, amino acids 581 and 582, amino acids 583 and 584, amino acids 584 and 585, amino acids 585 and 586, amino acids 586 and 587, amino acids 587 and 588, amino acids 588 and 589, or amino acids 589 and 590.
  • the insertion site can be between amino acids 575 and 576, amino acids 576 and 577, amino acids 577 and 578, amino acids 578 and 579, or amino acids 579 and 580.
  • the insertion site can be between amino acids 590 and 591, amino acids 591 and 592, amino acids 592 and 593, amino acids 593 and 594, amino acids 594 and 595, amino acids 595 and 596, amino acids 596 and 597, amino acids 597 and 598, amino acids 598 and 599, or amino acids 599 and 600.
  • the insertion site can be between amino acids 587 and 588 of AAV2, between amino acids 590 and 591 of AAV1, between amino acids 575 and 576 of AAV5, between amino acids 590 and 591 of AAV6, between amino acids 589 and 590 of AAV7, between amino acids 590 and 591 of AAV8, between amino acids 588 and 589 of AAV9, or between amino acids 589 and 590 of AAV10.
  • a subject rAAV virion comprises a peptide of from about 5 amino acids to about 11 amino acids in length inserted into the GH loop of the AAV capsid.
  • the insertion peptide has a length of 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, or 11 amino acids.
  • the insertion peptide can comprise an amino acid sequence of any one of the formulas set forth below.
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula I: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IIa: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IIb: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula III: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IV: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • the insertion peptide can comprise an amino acid sequence selected from LGETTRP (SEQ ID NO: 13), NETITRP (SEQ ID NO:14), KAGQANN (SEQ ID NO:15), KDPKTTN (SEQ ID NO:16), KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60).
  • LGETTRP SEQ ID NO: 13
  • NETITRP SEQ ID NO:14
  • KAGQANN SEQ ID NO:15
  • KDPKTTN SEQ ID NO:16
  • KDTDTTR SEQ ID NO:57
  • RAGGSVG SEQ ID NO:58
  • AVDTTKF SEQ ID NO:59
  • STGKVPN STGKVPN
  • the insertion peptide has from 1 to 4 spacer amino acids (Y 1 -Y 4 ) at the amino terminus and/or at the carboxyl terminus of any one of LGETTRP (SEQ ID NO: 13), NETITRP (SEQ ID NO:14), KAGQANN (SEQ ID NO:15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60).
  • Suitable spacer amino acids include, but are not limited to, leucine, alanine, glycine, and serine.
  • an insertion peptide has one of the following amino acid sequences: LALGETTRPA (SEQ ID NO:45); LANETITRPA (SEQ ID NO:46), LAKAGQANNA (SEQ ID NO:47), LAKDPKTTNA (SEQ ID NO:48), LAKDTDTTRA (SEQ ID NO:61), LARAGGSVGA (SEQ ID NO:62), LAAVDTTKFA (SEQ ID NO:63), and LASTGKVPNA (SEQ ID NO:64).
  • an insertion peptide has one of the following amino acid sequences: AALGETTRPA (SEQ ID NO:49); AANETITRPA (SEQ ID NO:50), AAKAGQANNA (SEQ ID NO:51), and AAKDPKTTNA (SEQ ID NO:52).
  • an insertion peptide has one of the following amino acid sequences: GLGETTRPA (SEQ ID NO:53); GNETITRPA (SEQ ID NO:54), GKAGQANNA (SEQ ID NO:55), and GKDPKTTNA (SEQ ID NO:56).
  • an insertion peptide comprises one of KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60), flanked on the C-terminus by AA and on the N-terminus by A; or comprises one of KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60) flanked on the C-terminus by G and on the N-terminus by A.
  • a subject rAAV virion capsid does not include any other amino acid substitutions, insertions, or deletions, other than an insertion of from about 7 amino acids to about 10 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject rAAV virion capsid includes from 1 to about 25 amino acid insertions, deletions, or substitutions, compared to the parental AAV capsid protein, in addition to an insertion of from about 7 amino acids to about 10 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject rAAV virion capsid includes from 1 to about 5, from about 5 to about 10, from about 10 to about 15, from about 15 to about 20, or from about 20 to about 25 amino acid insertions, deletions, or substitutions, compared to the parental AAV capsid protein, in addition to an insertion of from about 7 amino acids to about 10 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject rAAV virion capsid does not include one, two, three, or four, of the following amino acid substitutions: Y273F, Y444F, Y500F, and Y730F.
  • a subject variant capsid polypeptide comprises, in addition to an insertion peptide as described above, one, two, three, or four, of the following amino acid substitutions: Y273F, Y444F, Y500F, and Y730F.
  • a subject rAAV virion capsid is a chimeric capsid, e.g., the capsid comprises a portion of an AAV capsid of a first AAV serotype and a portion of an AAV capsid of a second serotype; and comprises an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject rAAV virion comprises a capsid protein comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 98%, or at least about 99%, amino acid sequence identity to the amino acid sequence provided in Figure 5 ; and an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein.
  • a subject rAAV virion comprises a capsid protein that includes a GH loop comprising an amino acid sequence having at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to an amino acid sequence set forth in Figure 18A-C .
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a retinal cell, compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a retinal cell, when administered via intravitreal injection, compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a photoreceptor (rod or cone) cell, compared to the infectivity of the photoreceptor cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a photoreceptor (rod or cone) cell, when administered via intravitreal injection, compared to the infectivity of the photoreceptor cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of an RGC, compared to the infectivity of the RGC by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of an RGC, when administered via intravitreal injection, compared to the infectivity of the RGC by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of an RPE cell, compared to the infectivity of the RPE cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of an RPE cell, when administered via intravitreal injection, compared to the infectivity of the RPE cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a Müller cell, compared to the infectivity of the Müller cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a Müller cell, when administered via intravitreal injection, compared to the infectivity of the Müller cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a bipolar cell, compared to the infectivity of the bipolar cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a bipolar cell, when administered via intravitreal injection, compared to the infectivity of the bipolar cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of an amacrine cell, compared to the infectivity of the amacrine cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of an amacrine cell, when administered via intravitreal injection, compared to the infectivity of the amacrine cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a horizontal cell, compared to the infectivity of the horizontal cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a horizontal cell, when administered via intravitreal injection, compared to the infectivity of the horizontal cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased ability to cross the internal limiting membrane (ILM), compared to the ability of an AAV virion comprising the corresponding parental AAV capsid protein to cross the ILM.
  • ILM internal limiting membrane
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased ability, when administered via intravitreal injection, to cross the internal limiting membrane (ILM), compared to the ability of an AAV virion comprising the corresponding parental AAV capsid protein to cross the ILM when administered via intravitreal injection.
  • ILM internal limiting membrane
  • a subject rAAV virion can cross the ILM, and can also traverse cell layers, including Müller cells, amacrine cells, etc., to reach the photoreceptor cells and or RPE cells.
  • a subject rAAV virion when administered via intravitreal injection, can cross the ILM, and can also traverse cell layers, including Müller cells, amacrine cells, etc., to reach the photoreceptor cells and or RPE cells.
  • a subject rAAV virion selectively infects a retinal cell, e.g., a subject rAAV virion infects a retinal cell with 10-fold, 15-fold, 20-fold, 25-fold, 50-fold, or more than 50-fold, specificity than a non-retinal cell, e.g., a cell outside the eye.
  • a subject rAAV virion selectively infects a retinal cell, e.g., a subject rAAV virion infects a photoreceptor cell with 10-fold, 15-fold, 20-fold, 25-fold, 50-fold, or more than 50-fold, specificity than a non-retinal cell, e.g., a cell outside the eye.
  • a retinal cell e.g., a subject rAAV virion infects a photoreceptor cell with 10-fold, 15-fold, 20-fold, 25-fold, 50-fold, or more than 50-fold, specificity than a non-retinal cell, e.g., a cell outside the eye.
  • a subject rAAV virion selectively infects a photoreceptor cell, e.g., a subject rAAV virion infects a photoreceptor cell with 10-fold, 15-fold, 20-fold, 25-fold, 50-fold, or more than 50-fold, specificity than a non-photoreceptor cell present in the eye, e.g., a retinal ganglion cell, a Müller cell, etc.
  • a subject rAAV virion exhibits at least 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, at least 50-fold, or more than 50-fold, increased infectivity of a photoreceptor cell, when administered via intravitreal injection, compared to the infectivity of the photoreceptor cell by an AAV virion comprising the corresponding parental AAV capsid protein, when administered via intravitreal injection.
  • a subject rAAV virion comprises a heterologous nucleic acid comprising a nucleotide sequence encoding a gene product.
  • the gene product is an interfering RNA.
  • the gene product is an aptamer.
  • the gene product is a polypeptide.
  • the gene product is a site-specific nuclease that provide for site-specific knock-down of gene function.
  • RNAi interfering RNA
  • suitable RNAi include RNAi that decrease the level of an apoptotic or angiogenic factor in a cell.
  • an RNAi can be an shRNA or siRNA that reduces the level of a gene product that induces or promotes apoptosis in a cell.
  • Genes whose gene products induce or promote apoptosis are referred to herein as "pro-apoptotic genes" and the products of those genes (mRNA; protein) are referred to as "pro-apoptotic gene products.”
  • Pro-apoptotic gene products include, e.g., Bax , Bid , Bak , and Bad gene products. See, e.g., U.S. Patent No. 7,846,730 .
  • Interfering RNAs could also be against an angiogenic product, for example VEGF (e.g., Cand5; see, e.g., U.S. Patent Publication No. 2011/0143400 ; U.S. Patent Publication No. 2008/0188437 ; and Reich et al. (2003) Mol. Vis. 9:210 ), VEGFR1 (e.g., Sirna-027; see, e.g., Kaiser et al. (2010) Am. J. Ophthalmol. 150:33 ; and Shen et al. (2006) Gene Ther. 13:225 ), or VEGFR2 ( Kou et al. (2005) Biochem. 44:15064 ). See also, U.S. Patent Nos.
  • VEGF e.g., Cand5; see, e.g., U.S. Patent Publication No. 2011/0143400 ; U.S. Patent Publication No. 2008/0188437 ; and Reich et al. (2003) Mol.
  • exemplary aptamers of interest include an aptamer against vascular endothelial growth factor (VEGF).
  • VEGF vascular endothelial growth factor
  • a VEGF aptamer can comprise the nucleotide sequence 5'-cgcaaucagugaaugcuuauacauccg-3' (SEQ ID NO:17).
  • a PDGF-specific aptamer e.g., E10030; see, e.g., Ni and Hui (2009) Ophthalmologica 223:401 ; and Akiyama et al. (2006) J. Cell Physiol. 207:407 ).
  • the polypeptide is generally a polypeptide that enhances function of a retinal cell, e.g., the function of a rod or cone photoreceptor cell, a retinal ganglion cell, a Müller cell, a bipolar cell, an amacrine cell, a horizontal cell, or a retinal pigmented epithelial cell.
  • polypeptides include neuroprotective polypeptides (e.g., GDNF, CNTF, NT4, NGF, and NTN); anti-angiogenic polypeptides (e.g., a soluble vascular endothelial growth factor (VEGF) receptor; a VEGF-binding antibody; a VEGF-binding antibody fragment (e.g., a single chain anti-VEGF antibody); endostatin; tumstatin; angiostatin; a soluble Flt polypeptide ( Lai et al. (2005) Mol. Ther. 12:659 ); an Fc fusion protein comprising a soluble Flt polypeptide (see, e.g., Pechan et al.
  • VEGF vascular endothelial growth factor
  • PEDF pigment epithelium-derived factor
  • TMP-3 tissue inhibitor of metalloproteinases-3
  • a light-responsive opsin e.g., a rhodopsin
  • anti-apoptotic polypeptides e.g., Bcl-2, Bcl-X1; and the like.
  • Suitable polypeptides include, but are not limited to, glial derived neurotrophic factor (GDNF); fibroblast growth factor 2; neurturin (NTN); ciliary neurotrophic factor (CNTF); nerve growth factor (NGF); neurotrophin-4 (NT4); brain derived neurotrophic factor (BDNF; e.g., a polypeptide comprising an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to a contiguous stretch of from about 200 amino acids to 247 amino acids of the amino acid sequence depicted in Figure 11 (SEQ ID NO:11)); epidermal growth factor; rhodopsin; X-linked inhibitor of apoptosis; and Sonic hedgehog.
  • GDNF glial derived neurotrophic factor
  • NTN neurturin
  • CNTF ciliary neurotrophic factor
  • NEF nerve growth factor
  • NT4 neurotrophin-4
  • BDNF
  • Suitable light-responsive opsins include, e.g., a light-responsive opsin as described in U.S. Patent Publication No. 2007/0261127 (e.g., ChR2; Chop2); U.S. Patent Publication No. 2001/0086421 ; U.S. Patent Publication No. 2010/0015095 ; and Diester et al. (2011) Nat. Neurosci. 14:387 .
  • Suitable polypeptides also include retinoschisin (e.g., a polypeptide comprising an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to a contiguous stretch of from about 200 amino acids to 224 amino acids of the amino acid sequence depicted in Figure 10 (SEQ ID NO:10).
  • Suitable polypeptides include, e.g., retinitis pigmentosa GTPase regulator (RGPR)-interacting protein-1 (see, e.g., GenBank Accession Nos.
  • Q96KN7, Q9EPQ2, and Q9GLM3 e.g., a polypeptide comprising an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to a contiguous stretch of from about 1150 amino acids to about 1200 amino acids, or from about 1200 amino acids to 1286 amino acids, of the amino acid sequence depicted in Figure 16 (SEQ ID NO:21); peripherin-2 (Prph2) (see, e.g., GenBank Accession No.
  • NP_000313 e.g., a polypeptide comprising an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to a contiguous stretch of from about 300 amino acids to 346 amino acids of the amino acid sequence depicted in Figure 14 (SEQ ID NO:19); and Travis et al.
  • peripherin e.g., a polypeptide comprising an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to a contiguous stretch of from about 400 amino acids to about 470 amino acids of the amino acid sequence depicted in Figure 15 (SEQ ID NO:20); a retinal pigment epithelium-specific protein (RPE65), (e.g., a polypeptide comprising an amino acid sequence having at least about 90%, at least about 95%, at least about 98%, at least about 99%, or 100%, amino acid sequence identity to a contiguous stretch of from about 200 amino acids to 247 amino acids of the amino acid sequence depicted in Figure 12 (SEQ ID NO:12)) (see, e.g., GenBank AAC39660; and Morimura et al. (1998) Proc. Natl. Acad. Sci. USA 95:3088 ); and the like.
  • RPE65 retinal pigment epithel
  • Suitable polypeptides also include: CHM (choroidermia (Rab escort protein 1)), a polypeptide that, when defective or missing, causes choroideremia (see, e.g., Donnelly et al. (1994) Hum. Mol. Genet. 3:1017 ; and van Bokhoven et al. (1994) Hum. Mol. Genet. 3:1041 ); and Crumbs homolog 1 (CRB1), a polypeptide that, when defective or missing, causes Leber congenital amaurosis and retinitis pigmentosa (see, e.g., den Hollander et al. (1999) Nat. Genet. 23:217 ; and GenBank Accession No. CAM23328).
  • CHM choroidermia (Rab escort protein 1)
  • a polypeptide that, when defective or missing, causes choroideremia see, e.g., Donnelly et al. (1994) Hum
  • Suitable polypeptides also include polypeptides that, when defective or missing, lead to achromotopsia, where such polypeptides include, e.g., cone photoreceptor cGMP-gated channel subunit alpha (CNGA3) (see, e.g., GenBank Accession No. NP_001289; and Booij et al. (2011) Ophthalmology 118:160-167 ); cone photoreceptor cGMP-gated cation channel beta-subunit (CNGB3) (see, e.g., Kohl et al.(2005) Eur J Hum Genet.
  • CNGA3 cone photoreceptor cGMP-gated channel subunit alpha
  • CNGB3 cone photoreceptor cGMP-gated cation channel beta-subunit
  • G protein guanine nucleotide binding protein
  • GNAT2 alpha transducing activity polypeptide 2
  • ACHM5 alpha transducing activity polypeptide 5
  • polypeptides that, when defective or lacking, lead to various forms of color blindness e.g., L-opsin, M-opsin, and S-opsin. See Mancuso et al. (2009) Nature 461(7265):784-787 .
  • a gene product of interest is a site-specific endonuclease that provide for site-specific knock-down of gene function, e.g., where the endonuclease knocks out an allele associated with a retinal disease.
  • a site-specific endonuclease can be targeted to the defective allele and knock out the defective allele.
  • a site-specific nuclease can also be used to stimulate homologous recombination with a donor DNA that encodes a functional copy of the protein encoded by the defective allele.
  • a subject rAAV virion can be used to deliver both a site-specific endonuclease that knocks out a defective allele, and can be used to deliver a functional copy of the defective allele, resulting in repair of the defective allele, thereby providing for production of a functional retinal protein (e.g., functional retinoschisin, functional RPE65, functional peripherin, etc.). See, e.g., Li et al.
  • a subject rAAV virion comprises a heterologous nucleotide sequence that encodes a site-specific endonuclease; and a heterologous nucleotide sequence that encodes a functional copy of a defective allele, where the functional copy encodes a functional retinal protein.
  • Functional retinal proteins include, e.g., retinoschisin, RPE65, retinitis pigmentosa GTPase regulator (RGPR)-interacting protein-1, peripherin, peripherin-2, and the like.
  • Site-specific endonucleases that are suitable for use include, e.g., zinc finger nucleases (ZFNs); and transcription activator-like effector nucleases (TALENs), where such site-specific endonucleases are non-naturally occurring and are modified to target a specific gene.
  • ZFNs zinc finger nucleases
  • TALENs transcription activator-like effector nucleases
  • site-specific endonucleases can be engineered to cut specific locations within a genome, and non-homologous end joining can then repair the break while inserting or deleting several nucleotides.
  • site-specific endonucleases also referred to as "INDELs” then throw the protein out of frame and effectively knock out the gene. See, e.g., U.S. Patent Publication No. 2011/0301073 .
  • a nucleotide sequence encoding a gene product of interest is operably linked to a constitutive promoter. In other embodiments, a nucleotide sequence encoding a gene product of interest is operably linked to an inducible promoter. In some instances, a nucleotide sequence encoding a gene product of interest is operably linked to a tissue-specific or cell type-specific regulatory element.
  • a nucleotide sequence encoding a gene product of interest is operably linked to a photoreceptor-specific regulatory element (e.g., a photoreceptor-specific promoter), e.g., a regulatory element that confers selective expression of the operably linked gene in a photoreceptor cell.
  • a photoreceptor-specific regulatory element e.g., a photoreceptor-specific promoter
  • Suitable photoreceptor-specific regulatory elements include, e.g., a rhodopsin promoter; a rhodopsin kinase promoter ( Young et al. (2003) Ophthalmol. Vis. Sci. 44:4076 ); a beta phosphodiesterase gene promoter ( Nicoud et al. (2007) J. Gene Med.
  • a retinitis pigmentosa gene promoter (Nicoud et al. (2007) supra ); an interphotoreceptor retinoid-binding protein (IRBP) gene enhancer (Nicoud et al. (2007) supra ); an IRBP gene promoter ( Yokoyama et al. (1992) Exp Eye Res. 55:225 ).
  • IRBP interphotoreceptor retinoid-binding protein
  • the present disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising: a) a subject rAAV virion, as described above; and b) a pharmaceutically acceptable carrier, diluent, excipient, or buffer.
  • the pharmaceutically acceptable carrier, diluent, excipient, or buffer is suitable for use in a human.
  • excipients include any pharmaceutical agent that can be administered without undue toxicity.
  • Pharmaceutically acceptable excipients include, but are not limited to, liquids such as water, saline, glycerol and ethanol.
  • Pharmaceutically acceptable salts can be included therein, for example, mineral acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, and the like; and the salts of organic acids such as acetates, propionates, malonates, benzoates, and the like.
  • auxiliary substances such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles.
  • the present disclosure provides a method of delivering a gene product to a retinal cell in an individual, the method comprising administering to the individual a subject rAAV virion as described above.
  • the gene product can be a polypeptide or an interfering RNA (e.g., an shRNA, an siRNA, and the like), an aptamer, or a site-specific endonuclease, as described above.
  • Delivering a gene product to a retinal cell can provide for treatment of a retinal disease.
  • the retinal cell can be a photoreceptor, a retinal ganglion cell, a Müller cell, a bipolar cell, an amacrine cell, a horizontal cell, or a retinal pigmented epithelial cell.
  • the retinal cell is a photoreceptor cell, e.g., a rod or cone cell.
  • the present disclosure provides a method of treating a retinal disease, the method comprising administering to an individual in need thereof an effective amount of a subject rAAV virion as described above.
  • a subject rAAV virion can be administered via intraocular injection, by intravitreal injection, or by any other convenient mode or route of administration.
  • Other convenient modes or routes of administration include, e.g., intravenous, intranasal, etc.
  • a “therapeutically effective amount” will fall in a relatively broad range that can be determined through experimentation and/or clinical trials.
  • a therapeutically effective dose will be on the order of from about 10 6 to about 10 15 of the rAAV virions, e.g., from about 10 8 to 10 12 rAAV virions.
  • an effective amount of rAAV virions to be delivered to cells will be on the order of from about 10 8 to about 10 13 of the rAAV virions.
  • Other effective dosages can be readily established by one of ordinary skill in the art through routine trials establishing dose response curves.
  • more than one administration may be employed to achieve the desired level of gene expression over a period of various intervals, e.g., daily, weekly, monthly, yearly, etc.
  • Ocular diseases that can be treated using a subject method include, but are not limited to, acute macular neuroretinopathy; Behcet's disease; choroidal neovascularization; diabetic uveitis; histoplasmosis; macular degeneration, such as acute macular degeneration, non-exudative age related macular degeneration and exudative age related macular degeneration; edema, such as macular edema, cystoid macular edema and diabetic macular edema; multifocal choroiditis; ocular trauma which affects a posterior ocular site or location; ocular tumors; retinal disorders, such as central retinal vein occlusion, diabetic retinopathy (including proliferative diabetic retinopathy), proliferative vitreoretinopathy (PVR), retinal arterial occlusive disease, retinal detachment, uveitic retinal disease; sympathetic opthalmia; Vogt Koyanagi-Ha
  • the present disclosure provides an isolated nucleic acid comprising a nucleotide sequence that encodes a subject variant adeno-associated virus (AAV) capsid protein as described above, where the variant AAV capsid protein comprises an insertion of from about 5 amino acids to about 11 amino acids in the GH loop or loop IV relative to a corresponding parental AAV capsid protein, and where the variant capsid protein, when present in an AAV virion, provides for increased infectivity of a retinal cell compared to the infectivity of the retinal cell by an AAV virion comprising the corresponding parental AAV capsid protein.
  • a subject isolated nucleic acid can be an AAV vector, e.g., a recombinant AAV vector.
  • a variant AAV capsid protein encoded by a subject nucleic acid has an insertion peptide of from about 5 amino acids to about 11 amino acids in length is inserted into the GH loop of an AAV capsid.
  • the insertion peptide has a length of 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, or 11 amino acids.
  • the insertion peptide can comprise an amino acid sequence of any one of the formulas set forth below.
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula I: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IIa: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IIb: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula III: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • an insertion peptide can be a peptide of from 5 to 11 amino acids in length, where the insertion peptide is of Formula IV: Y 1 Y 2 X 1 X 2 X 3 X 4 X 5 X 6 X 7 Y 3 Y 4 where:
  • the insertion peptide can comprise an amino acid sequence selected from LGETTRP (SEQ ID NO: 13), NETITRP (SEQ ID NO:14), KAGQANN (SEQ ID NO:15), KDPKTTN (SEQ ID NO:16), KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60).
  • LGETTRP SEQ ID NO: 13
  • NETITRP SEQ ID NO:14
  • KAGQANN SEQ ID NO:15
  • KDPKTTN SEQ ID NO:16
  • KDTDTTR SEQ ID NO:57
  • RAGGSVG SEQ ID NO:58
  • AVDTTKF SEQ ID NO:59
  • STGKVPN STGKVPN
  • the insertion peptide has from 1 to 4 spacer amino acids (Y 1 -Y 4 ) at the amino terminus and/or at the carboxyl terminus of any one of LGETTRP (SEQ ID NO: 13), NETITRP (SEQ ID NO:14), KAGQANN (SEQ ID NO:15), KDPKTTN (SEQ ID NO: 16), KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60).
  • Suitable spacer amino acids include, but are not limited to, leucine, alanine, glycine, and serine.
  • an insertion peptide has one of the following amino acid sequences: LALGETTRPA (SEQ ID NO:45); LANETITRPA (SEQ ID NO:46), LAKAGQANNA (SEQ ID NO:47), LAKDPKTTNA (SEQ ID NO:48), LAKDTDTTRA (SEQ ID NO:61), LARAGGSVGA (SEQ ID NO:62), LAAVDTTKFA (SEQ ID NO:63), and LASTGKVPNA (SEQ ID NO:64).
  • an insertion peptide has one of the following amino acid sequences: AALGETTRPA (SEQ ID NO:49); AANETITRPA (SEQ ID NO:50), AAKAGQANNA (SEQ ID NO:51), and AAKDPKTTNA (SEQ ID NO:52).
  • an insertion peptide has one of the following amino acid sequences: GLGETTRPA (SEQ ID NO:53); GNETITRPA (SEQ ID NO:54), GKAGQANNA (SEQ ID NO:55), and GKDPKTTNA (SEQ ID NO:56).
  • an insertion peptide comprises one of KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60), flanked on the C-terminus by AA and on the N-terminus by A; or comprises one of KDTDTTR (SEQ ID NO:57), RAGGSVG (SEQ ID NO:58), AVDTTKF (SEQ ID NO:59), and STGKVPN (SEQ ID NO:60) flanked on the C-terminus by G and on the N-terminus by A.
  • a subject recombinant AAV vector can be used to generate a subject recombinant AAV virion, as described above.
  • the present disclosure provides a recombinant AAV vector that, when introduced into a suitable cell, can provide for production of a subject recombinant AAV virion.
  • the present invention further provides host cells, e.g., isolated (genetically modified) host cells, comprising a subject nucleic acid.
  • a subject host cell can be an isolated cell, e.g., a cell in in vitro culture.
  • a subject host cell is useful for producing a subject rAAV virion, as described below. Where a subject host cell is used to produce a subject rAAV virion, it is referred to as a "packaging cell.”
  • a subject host cell is stably genetically modified with a subject nucleic acid.
  • a subject host cell is transiently genetically modified with a subject nucleic acid.
  • a subject nucleic acid is introduced stably or transiently into a host cell, using established techniques, including, but not limited to, electroporation, calcium phosphate precipitation, liposome-mediated transfection, and the like.
  • a subject nucleic acid will generally further include a selectable marker, e.g., any of several well-known selectable markers such as neomycin resistance, and the like.
  • a subject host cell is generated by introducing a subject nucleic acid into any of a variety of cells, e.g., mammalian cells, including, e.g., murine cells, and primate cells (e.g., human cells).
  • Suitable mammalian cells include, but are not limited to, primary cells and cell lines, where suitable cell lines include, but are not limited to, 293 cells, COS cells, HeLa cells, Vero cells, 3T3 mouse fibroblasts, C3H10T1/2 fibroblasts, CHO cells, and the like.
  • suitable host cells include, e.g., HeLa cells (e.g., American Type Culture Collection (ATCC) No. CCL-2), CHO cells (e.g., ATCC Nos.
  • CRL9618, CCL61, CRL9096 293 cells (e.g., ATCC No. CRL-1573), Vero cells, NIH 3T3 cells (e.g., ATCC No. CRL-1658), Huh-7 cells, BHK cells (e.g., ATCC No. CCL10), PC12 cells (ATCC No. CRL1721), COS cells, COS-7 cells (ATCC No. CRL1651), RAT1 cells, mouse L cells (ATCC No. CCLI.3), human embryonic kidney (HEK) cells (ATCC No. CRL1573), HLHepG2 cells, and the like.
  • a subject host cell can also be made using a baculovirus to infect insect cells such as Sf9 cells, which produce AAV (see, e.g., U.S. Patent No. 7,271,002 ; US patent application 12/297,958 )
  • a subject genetically modified host cell includes, in addition to a nucleic acid comprising a nucleotide sequence encoding a variant AAV capsid protein, as described above, a nucleic acid that comprises a nucleotide sequence encoding one or more AAV rep proteins.
  • a subject host cell further comprises an rAAV vector.
  • An rAAV virion can be generated using a subject host cell. Methods of generating an rAAV virion are described in, e.g., U.S. Patent Publication No. 2005/0053922 and U.S. Patent Publication No. 2009/0202490 .
  • Standard abbreviations may be used, e.g., bp, base pair(s); kb, kilobase(s); pl, picoliter(s); s or sec, second(s); min, minute(s); h or hr, hour(s); aa, amino acid(s); kb, kilobase(s); bp, base pair(s); nt, nucleotide(s); i.m., intramuscular(ly); i.p., intraperitoneal(ly); s.c., subcutaneous(ly); and the like.
  • Example 1 AAV variant with enhanced transduction of retinal cells
  • the approach used was to create a peptide display library by introducing a unique Avr II site into the wild type AAV2 genome between amino acid 587 and 588 by polymerase chain reaction (PCR) mutagenesis.
  • a random 21 nucleotide insert, 7mer For was used to synthesize dsDNA inserts, along with the antisense primer 7mer Rev.
  • the resulting dsDNA inserts were cloned into the Avr II site of the genome after digestion with Nhe I, producing a diverse 7mer display library which was then packaged (Perabo et al., 2003; Muller et al., 2003).
  • the virus was generated such that each viral genome was packaged or encapsidated within the capsid protein variant that that genome encoded. In this respect, functional improvements identified through selection can be linked to the genome sequence encoding this improved function contained within the viral capsid.
  • This library was subjected to positive selection within rho-GFP mice ( Wensel et al. (2005) Vision Res. 45:3445 ). Briefly, in one round of selection, adult rho-GFP mice were intravitreally injected with 2 ⁇ L of phosphate buffered saline (PBS)-dialyzed, iodixanol-purified library with a genomic titer of approximately 1x10 12 viral genomes (vg)/mL. An ultrafine 30 1/2-gauge disposable needle was passed through the sclera of the animal's eye, at the equator and next to the limbus, into the vitreous cavity.
  • PBS phosphate buffered saline
  • Injection of 2 ⁇ l of virus was made with direct observation of the needle in the center of the vitreous cavity.
  • One week post-injection eyes were enucleated and retinas dissociated using a light, papain protease treatment, followed by fluorescence activated cell sorter (FACS) isolation of photoreceptor populations.
  • FACS fluorescence activated cell sorter
  • cap genes of fifty variants were sequenced to determine the most prominent and successful variants to have permissive mutations for intravitreal photoreceptor transduction.
  • 46 gave readable sequences of a 7mer insert.
  • nearly two thirds of clones contained the same distinct 7mer motif ( ⁇ 588 LGETTRP ⁇ ; SEQ ID NO: 13).
  • the next most prominent variant ⁇ 588 NETITRP ⁇ ; SEQ ID NO: 14
  • TRP nonpolar proline residue
  • Frequency (%) Frequency ⁇ 588 LGETTRP ⁇ (SEQ ID NO:13) 64 31 ⁇ 588 NETITRP ⁇ (SEQ ID NO:14) 12 5 ⁇ 588 KAGQANN ⁇ (SEQ ID NO:15) 6 3 ⁇ 588 KDPKTTN ⁇ (SEQ ID NO:16) 4 2 ⁇ 588 KDTDTTR (SEQ ID NO:57) 2 ⁇ 588 RAGGSVG (SEQ ID NO:58) 1 ⁇ 588 AVDTTKF (SEQ ID NO:59) 1 ⁇ 588 STGKVPN (SEQ ID NO:60) 1
  • FIG. 1 Representative three-dimensional capsid model of AAV2 containing a random heptamer (shown in orange) following amino acid 587. This area of the AAV2 capsid likely participates in cell-surface receptor binding.
  • FIG. 2 AAV2 7M8 variant (right) demonstrates greater levels of intravitreal photoreceptor transduction relative to AAV2 (left). Confocal microscopy of transverse retinal sections three weeks after intravitreal injection of 2 ⁇ L of 1x10 12 vg/mL of AAV2 7M8 and AAV2 scCAG GFP in adult mice. Red arrows (top) denote photoreceptor nuclei and blue arrow (top) denote photoreceptor outer segments.
  • FIG. 3 Representative fluorescence images of retinal cryoslices showing GFP expression resulting from 7m8 carrying the GFP gene under the control of the ubiquitous CAG promoter (left) or a photoreceptor specific Rho promoter (right).
  • FIG. 4 GFP(+) photoreceptor cells per million retinal cells as counted by flow cytometry. 7m8 transduces more than 2x the amount of photoreceptors compared 7m8 bearing 4 tyrosine mutations (top).
  • a functional retinoschisin (RS1) protein was delivered to retinoschisin-deficient mice (Rs1h-deficient mice; Rs1h is the mouse homolog of human RS1).
  • the vector comprises a nucleotide sequence encoding a functional retinoschisin protein under transcriptional control of a rhodopsin promoter. See Figures 13A-C , where the bold and underlined nucleotide sequence (nucleotides 4013-4851) are the rhodopsin promoter; and nucleotides 4866-5540 (with the start atg and stop tga sequences shown in bold) encode a human RS1 protein.
  • the 7m8-rho-RS1 construct was administered intravitreally to Rslh-/- mice at P15.
  • Rslh-/- mice were generated through targeted disruption of exon 3 of the Rs1h gene, as described ( Weber et al. (2002) Proc. Natl. Acad. Sci. USA 99:6222 ).
  • the Rslh-/- mice are deficient in the Rs1h protein product, have an electronegative ERG (e.g., a reduced b-wave with relative preservation of the a-wave) and splitting of the layers of the retina, similar to what is seen in human retinoschisis patients.
  • ERG electronegative ERG
  • FIGS 7A-I Representative high-resolution SD-OCT images of retinas injected with 7m8-rho-GFP (left column), 7m8-rho-RS1 (middle column), or uninjected WT animals (right column). Fundus images were taken through the inner nuclear layer of the superior retina and exclude other layers (a-c). Transverse images of the superior (d-f) and inferior (g-i) retina were taken using the optic nerve head as a landmark.
  • the untreated RS1 retina increases in overall thickness when measured from the inner limiting membrane (ILM) to the photoreceptors, as the pathology progresses due to the schisis splitting the inner retina. This process is distinct from that observed in most retinal degenerative diseases (RDD) which do not form schisis, but exhibit progressive photoreceptor cell death in the INL and concomitant retinal thinning and loss of ERG amplitude.
  • RDD retinal degenerative diseases
  • the ONL thins as photoreceptors die from the disease, but this is distinct from the overall retinal thickness change. It is generally thought that a successful therapy for RS1 would return the overall retinal thickness to the wildtype and ameliorate the photoreceptor loss in the ONL.
  • RS1 is one of the very few examples of a retinal disease in which the pathology increases the retinal thickness with concomitant erg amplitude loss.
  • an extracellular retinal "glue; - thins the retina back to the wildtype thickness and the erg amplitude returns to near normal levels as the schisis resolves.
  • Figure 8a shows a comparison of functional rescue of untreated Rs1-/- eyes to AAV2-rho-RS1, 7m8-rho-GFP, and 7m8-rho-RS1 injected eyes both one month (left) and 4 months (right) after injection.
  • 7m8-rho-RS1 led to considerable rescue of the ERG b-wave amplitude, whereas AAV2-rho.RS1 was statistically indistinguishable from untreated eyes.
  • Figure 8b shows representative ERG traces from 7m8-rho-RS1-injected eyes show improved amplitude of the a-wave and b-wave and a waveform closer to wild-type eyes, compared to 7m8-rho-GFP-injected eyes.
  • Figure 8c shows the amplitude of the full-field scotopic b-wave resulting from a high intensity (1 log cd x s/m2) stimulus was recorded on a monthly basis beginning one month after injection at P15 for each condition. Three responses were recorded and averaged for each eye at each time point.
  • Figures 9A-E Sustained improvements in retinal thickness measured at 10 months post 7m8-rho-RS1 treatment.
  • Measurements of c) retinal thickness, d) ONL thickness, and e) and inner and outer segment thickness are plotted as a function of distance from the optic nerve head.
  • Example 3 AAV variant used to deliver a protein to retinal cells in the macaque
  • a recombinant AAV2 virion (7m8 carrying GFP under the control of a connexin36 promoter) was generated.
  • the recombinant AAV2 virion included an AAV2 capsid variant with an insertion of LALGETTRPA peptide between amino acids 587 and 588 of AAV2 capsid, and GFP under transcriptional control of a connexin36 promoter, which is expressed in interneurons.
  • the rAAV2 virion was injected intravitreally into the eye of a macaque. The data are shown in Figure 18 .
  • Figure 18 provides a fluorescence fundus image showing GFP expression at the back of the retina 9 weeks after administration of 7m8 carrying GFP under the control of a connexin36 promoter. Compared to the parental AAV2 serotype ( Yin et al, IOVS 52(5); 2775 ), a higher level of expression was seen in the foveal ring, and visible fluorescence was seen in the central retina outside the fovea.

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Families Citing this family (230)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9233131B2 (en) 2003-06-30 2016-01-12 The Regents Of The University Of California Mutant adeno-associated virus virions and methods of use thereof
US9441244B2 (en) 2003-06-30 2016-09-13 The Regents Of The University Of California Mutant adeno-associated virus virions and methods of use thereof
US8663624B2 (en) 2010-10-06 2014-03-04 The Regents Of The University Of California Adeno-associated virus virions with variant capsid and methods of use thereof
KR20220056884A (ko) 2011-04-22 2022-05-06 더 리젠츠 오브 더 유니버시티 오브 캘리포니아 변이체 캡시드를 지니는 아데노-관련 바이러스 및 이의 사용 방법
TWI702955B (zh) 2012-05-15 2020-09-01 澳大利亞商艾佛蘭屈澳洲私營有限公司 使用腺相關病毒（ａａｖ）ｓｆｌｔ-１治療老年性黃斑部退化（ａｍｄ）
ES2883590T3 (es) 2012-12-12 2021-12-09 Broad Inst Inc Suministro, modificación y optimización de sistemas, métodos y composiciones para la manipulación de secuencias y aplicaciones terapéuticas
ES2701749T3 (es) 2012-12-12 2019-02-25 Broad Inst Inc Métodos, modelos, sistemas y aparatos para identificar secuencias diana para enzimas Cas o sistemas CRISPR-Cas para secuencias diana y transmitir resultados de los mismos
US10266845B2 (en) 2013-02-08 2019-04-23 The Trustees Of The University Of Pennsylvania Enhanced AAV-mediated gene transfer for retinal therapies
US9567376B2 (en) 2013-02-08 2017-02-14 The Trustees Of The University Of Pennsylvania Enhanced AAV-mediated gene transfer for retinal therapies
KR102346455B1 (ko) 2013-03-15 2022-01-04 더 트러스티스 오브 더 유니버시티 오브 펜실베니아 Ｍｐｓｉ 치료를 위한 조성물 및 방법
AU2014265799B2 (en) * 2013-05-16 2019-10-31 Applied Genetic Technologies Corporation Promoters, expression cassettes, vectors, kits, and methods for the treatment of achromatopsia and other diseases
ES2897508T3 (es) 2013-05-31 2022-03-01 Univ California Variantes de virus adenoasociados y métodos de uso de las mismas
EP3674411A1 (de) 2013-06-17 2020-07-01 The Broad Institute, Inc. Bereitstellung, technische konzeption und optimierung von tandemführungssystemen, verfahren und zusammensetzungen zur sequenzmanipulation
AU2014281027A1 (en) 2013-06-17 2016-01-28 Massachusetts Institute Of Technology Optimized CRISPR-Cas double nickase systems, methods and compositions for sequence manipulation
EP3011033B1 (de) 2013-06-17 2020-02-19 The Broad Institute, Inc. Funktionale genomik unter verwendung von crispr-cas-systemen, zusammensetzungen, verfahren, schirme und anwendungen davon
CA2915842C (en) 2013-06-17 2022-11-29 The Broad Institute, Inc. Delivery and use of the crispr-cas systems, vectors and compositions for hepatic targeting and therapy
RU2716421C2 (ru) 2013-06-17 2020-03-11 Те Брод Инститьют Инк. Доставка, применение и применения в терапии систем crispr-cas и композиций для целенаправленного воздействия на нарушения и заболевания с использованием вирусных компонентов
WO2014207190A1 (en) * 2013-06-28 2014-12-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for expressing a polynucleotide of interest in the retina of a subject
EP3702461A1 (de) 2013-07-08 2020-09-02 Institut National De La Sante Et De La Recherche Medicale - Inserm Verfahren zur durchführung von oligonucleotidvermitteltem antisense-exon-skipping in der netzhaut einer darauf angewiesenen person
EP3561062A1 (de) 2013-09-13 2019-10-30 California Institute of Technology Selektive rückgewinnung
WO2015089364A1 (en) 2013-12-12 2015-06-18 The Broad Institute Inc. Crystal structure of a crispr-cas system, and uses thereof
WO2015089486A2 (en) 2013-12-12 2015-06-18 The Broad Institute Inc. Systems, methods and compositions for sequence manipulation with optimized functional crispr-cas systems
SG10201804976YA (en) * 2013-12-12 2018-07-30 Broad Inst Inc Delivery, Use and Therapeutic Applications of the Crispr-Cas Systems and Compositions for Genome Editing
WO2015089351A1 (en) 2013-12-12 2015-06-18 The Broad Institute Inc. Compositions and methods of use of crispr-cas systems in nucleotide repeat disorders
GB201403684D0 (en) * 2014-03-03 2014-04-16 King S College London Vector
WO2015126972A1 (en) * 2014-02-21 2015-08-27 University Of Florida Research Foundation, Inc. Methods and compositions for gene delivery to on bipolar cells
CA2941640A1 (en) 2014-03-04 2015-09-11 University Of Florida Research Foundation, Inc. Improved raav vectors and methods for transduction of photoreceptors and rpe cells
EP3957735A1 (de) 2014-03-05 2022-02-23 Editas Medicine, Inc. Verfahren im zusammenhang mit crispr/cas und verfahren zur behandlung des usher-syndroms und von retinitis pigmentosa
US11141493B2 (en) 2014-03-10 2021-10-12 Editas Medicine, Inc. Compositions and methods for treating CEP290-associated disease
US11339437B2 (en) 2014-03-10 2022-05-24 Editas Medicine, Inc. Compositions and methods for treating CEP290-associated disease
DK3116997T3 (da) * 2014-03-10 2019-08-19 Editas Medicine Inc Crispr/cas-relaterede fremgangsmåder og sammensætninger til behandling af lebers kongenitale amaurose 10 (lca10)
EP3800191A1 (de) 2014-03-17 2021-04-07 Adverum Biotechnologies, Inc. Zusammensetzungen und verfahren zur verbesserten genexpression in zapfenzellen
EP3981876A1 (de) 2014-03-26 2022-04-13 Editas Medicine, Inc. Verfahren und zusammensetzungen in zusammenhang mit crispr/cas zur behandlung von sichelzellanämie
AU2015252797C1 (en) * 2014-05-02 2021-04-22 Genzyme Corporation AAV vectors for retinal and CNS gene therapy
WO2015191508A1 (en) 2014-06-09 2015-12-17 Voyager Therapeutics, Inc. Chimeric capsids
WO2016018816A1 (en) * 2014-07-31 2016-02-04 The Board Of Regents Of The University Of Oklahoma High isomerohydrolase activity mutants of mammalian rpe65
MX2017003426A (es) * 2014-09-16 2017-07-28 Genzyme Corp Vectores virales adeno-asociados para tratar glaucoma de miocilina (myoc).
JP6401871B2 (ja) 2014-11-05 2018-10-10 ボイジャー セラピューティクス インコーポレイテッドＶｏｙａｇｅｒ Ｔｈｅｒａｐｅｕｔｉｃｓ，Ｉｎｃ． パーキンソン病の治療のためのａａｄｃポリヌクレオチド
KR102584655B1 (ko) 2014-11-14 2023-10-06 보이저 테라퓨틱스, 인크. 조절성 폴리뉴클레오티드
KR20230169197A (ko) 2014-11-14 2023-12-15 보이저 테라퓨틱스, 인크. 근위축성 측삭 경화증(als)을 치료하는 조성물 및 방법
EP3889260A1 (de) 2014-12-12 2021-10-06 The Broad Institute, Inc. Geschützte guide-rnas (pgrnas)
US11697825B2 (en) 2014-12-12 2023-07-11 Voyager Therapeutics, Inc. Compositions and methods for the production of scAAV
US10584321B2 (en) * 2015-02-13 2020-03-10 University Of Massachusetts Compositions and methods for transient delivery of nucleases
US10648000B2 (en) 2015-02-16 2020-05-12 University Of Florida Research Foundation, Incorporated rAAV vector compositions, methods for targeting vascular endothelial cells and use in treatment of type I diabetes
WO2016134375A1 (en) * 2015-02-20 2016-08-25 University Of Iowa Research Foundation Methods and compositions for treating genetic eye diseases
AU2016226289B2 (en) * 2015-03-02 2021-04-29 Adverum Biotechnologies, Inc. Compositions and methods for intravitreal delivery of polynucleotides to retinal cones
EP3277819B1 (de) * 2015-03-24 2021-03-03 The Regents of The University of California Adenoassoziierte virenvarianten und verfahren zur verwendung davon
US10081659B2 (en) 2015-04-06 2018-09-25 The United States Of America, As Represented By The Secretary, Dept. Of Health And Human Services Adeno-associated vectors for enhanced transduction and reduced immunogenicity
US11180793B2 (en) 2015-04-24 2021-11-23 Editas Medicine, Inc. Evaluation of Cas9 molecule/guide RNA molecule complexes
EP3294332A4 (de) * 2015-05-14 2018-12-26 Joslin Diabetes Center, Inc. Retinolbindendes protein 3 (rbp3) als schutzfaktor bei nicht diabetischer netzhautdegeneration
CN109536474A (zh) 2015-06-18 2019-03-29 布罗德研究所有限公司 降低脱靶效应的crispr酶突变
WO2016205759A1 (en) 2015-06-18 2016-12-22 The Broad Institute Inc. Engineering and optimization of systems, methods, enzymes and guide scaffolds of cas9 orthologs and variants for sequence manipulation
AU2016363830C1 (en) * 2015-12-04 2022-03-24 Centre National De La Recherche Scientifique Promoters and uses thereof
BR112018011711A2 (pt) * 2015-12-11 2018-12-04 California Inst Of Techn peptídeos de direcionamento para direcionar virus adeno-associados (aav)
WO2017099838A1 (en) 2015-12-11 2017-06-15 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Gene therapy for combined methylmalonic acidemia/aciduria and hyperhomocysteinemia/homocystinuria, cobalamin c type, and deficiency of mmachc
GB2545763A (en) 2015-12-23 2017-06-28 Adverum Biotechnologies Inc Mutant viral capsid libraries and related systems and methods
KR20180121899A (ko) 2016-02-03 2018-11-09 더 트러스티스 오브 더 유니버시티 오브 펜실바니아 I형 점액다당류증을 치료하기 위한 유전자 요법
US11512311B2 (en) 2016-03-25 2022-11-29 Editas Medicine, Inc. Systems and methods for treating alpha 1-antitrypsin (A1AT) deficiency
AU2017248731A1 (en) 2016-04-15 2018-10-18 The Trustees Of The University Of Pennsylvania Compositions for treatment of wet age-related macular degeneration
CN110267673B (zh) 2016-04-29 2023-05-16 珍视生物股份公司 利用chrimson来进行光遗传视觉恢复
WO2017189964A2 (en) 2016-04-29 2017-11-02 Voyager Therapeutics, Inc. Compositions for the treatment of disease
WO2017189959A1 (en) 2016-04-29 2017-11-02 Voyager Therapeutics, Inc. Compositions for the treatment of disease
WO2017190125A1 (en) 2016-04-29 2017-11-02 Adverum Biotechnologies, Inc. Evasion of neutralizing antibodies by a recombinant adeno-associated virus
WO2017192750A1 (en) * 2016-05-04 2017-11-09 Oregon Health & Science University Recombinant adeno-associated viral vectors
CN109476707B (zh) * 2016-05-13 2022-12-02 4D分子治疗有限公司 腺相关病毒变体衣壳和其使用方法
SG11201809699XA (en) 2016-05-18 2018-12-28 Voyager Therapeutics Inc Modulatory polynucleotides
WO2017201258A1 (en) 2016-05-18 2017-11-23 Voyager Therapeutics, Inc. Compositions and methods of treating huntington's disease
MA44546B1 (fr) * 2016-06-15 2021-03-31 Univ California Virus adéno-associés variants et procédés d'utilisation
KR20210021113A (ko) * 2016-06-16 2021-02-24 애드베룸 바이오테크놀로지스, 인코포레이티드 Aav2 변이체를 아플리베르셉트와 함께 사용하는 amd의 치료
DK3472317T3 (da) * 2016-06-16 2022-05-23 Adverum Biotechnologies Inc Sammensætninger og fremgangsmåder til reduktion af okulær vaskularisation
BR112019001815A2 (pt) * 2016-07-29 2019-05-07 The Regents Of The University Of California vírions do vírus adeno-associado com capsídeo variante e seus métodos de uso
BR112019001887A2 (pt) 2016-08-02 2019-07-09 Editas Medicine Inc composições e métodos para o tratamento de doença associada a cep290
CN110650673B (zh) 2016-08-30 2024-04-09 加利福尼亚大学董事会 用于生物医学靶向和递送的方法以及用于实践该方法的装置和系统
US11192925B2 (en) 2016-10-19 2021-12-07 Adverum Biotechnologies, Inc. Modified AAV capsids and uses thereof
AU2017372351B2 (en) * 2016-11-06 2019-12-12 Nanoscope Technologies Llc Optogenetic modulation by multi-characteristic opsins for vision restoration and other applications thereof
US11236360B2 (en) 2016-12-09 2022-02-01 Regents Of The University Of Minnesota Adeno-associated viruses engineered for selectable tropism
CA3054687A1 (en) * 2017-02-28 2018-09-07 Adverum Biotechnologies, Inc. Modified aav capsids and uses thereof
EP3596217A1 (de) 2017-03-14 2020-01-22 Editas Medicine, Inc. Systeme und verfahren zur behandlung von hämoglobinopathien
EA201992001A1 (ru) 2017-03-17 2020-01-24 Адверум Байотекнолоджис, Инк. Композиции и способы повышения экспрессии генов
EP3609542B1 (de) * 2017-04-11 2023-02-22 Ruprecht-Karls-Universität Heidelberg Adeno-assoziierte virusbibliothek
JP2020516291A (ja) 2017-04-14 2020-06-11 レジェンクスバイオ インコーポレーテッド ヒト神経細胞又はグリア細胞により生産された組換えヒトイズロン酸−２−スルファターゼ（ｉｄｓ）によるムコ多糖症ｉｉ型の治療
EP3619308A4 (de) 2017-05-05 2021-01-27 Voyager Therapeutics, Inc. Zusammensetzungen und verfahren zur behandlung von morbus huntington
AU2018261790A1 (en) 2017-05-05 2019-11-28 Voyager Therapeutics, Inc. Compositions and methods of treating amyotrophic lateral sclerosis (ALS)
EP3622070A2 (de) 2017-05-10 2020-03-18 Editas Medicine, Inc. Crispr/rna-geführte nukleasesysteme und -verfahren
JOP20190269A1 (ar) 2017-06-15 2019-11-20 Voyager Therapeutics Inc بولي نوكليوتيدات aadc لعلاج مرض باركنسون
AU2018290954B2 (en) * 2017-06-30 2023-07-13 The Regents Of The University Of California Adeno-associated virus virions with variant capsids and methods of use thereof
KR20240063170A (ko) 2017-07-06 2024-05-09 더 트러스티스 오브 더 유니버시티 오브 펜실베니아 I형 점액다당류증을 치료하기 위한 유전자 요법
CN111132626B (zh) 2017-07-17 2024-01-30 沃雅戈治疗公司 轨迹阵列引导系统
US11680249B2 (en) 2017-08-28 2023-06-20 The Regents Of The University Of California Adeno-associated virus capsid variants and methods of use thereof
US11118192B2 (en) 2017-09-20 2021-09-14 4D Molecular Therapeutics Inc. Adeno-associated virus variant capsids and methods of use thereof
US20200308553A1 (en) * 2017-10-16 2020-10-01 Vigeneron Gmbh Aav vectors
US11434502B2 (en) 2017-10-16 2022-09-06 Voyager Therapeutics, Inc. Treatment of amyotrophic lateral sclerosis (ALS)
US20200237799A1 (en) 2017-10-16 2020-07-30 Voyager Therapeutics, Inc. Treatment of amyotrophic lateral sclerosis (als)
AU2018350992A1 (en) 2017-10-18 2020-05-21 Regenxbio Inc. Fully-human post-translationally modified antibody therapeutics
EP3697449A1 (de) 2017-10-18 2020-08-26 REGENXBIO Inc. Behandlung von augenkrankheiten und metastasierendem dickdarmkrebs mit menschlicher post-translational modifizierter vegf-falle
WO2019077159A1 (en) * 2017-10-20 2019-04-25 INSERM (Institut National de la Santé et de la Recherche Médicale) METHODS OF EXPRESSING POLYNUCLEOTIDE OF INTEREST IN CONE PHOTORECCEPTORS OF A SUBJECT COMPRISING SUB-RETINAL ADMINISTRATION OF A THERAPEUTICALLY EFFECTIVE AMOUNT OF A RECOMBINANT AAV9 DERIVATIVE VECTOR
US11851671B2 (en) 2017-10-23 2023-12-26 Regents Of The University Of Minnesota Programmable assembly of virus composites for receptor-targeted gene delivery
FI3717636T3 (fi) 2017-11-27 2023-06-01 4D Molecular Therapeutics Inc Adenoassosioidun viruksen kapsidivariantteja ja käyttö angiogeneesin estämisessä
US10610606B2 (en) 2018-02-01 2020-04-07 Homology Medicines, Inc. Adeno-associated virus compositions for PAH gene transfer and methods of use thereof
EP3749754A4 (de) 2018-02-08 2021-10-20 Applied Stemcell, Inc. Verfahren für das screening von varianten eines zielgens
US11306329B2 (en) 2018-02-19 2022-04-19 City Of Hope Adeno-associated virus compositions for restoring F8 gene function and methods of use thereof
AU2019247191A1 (en) * 2018-04-03 2020-10-15 Ginkgo Bioworks, Inc. Virus vectors for targeting ophthalmic tissues
US20210231560A1 (en) 2018-04-29 2021-07-29 Regenxbio Inc. Systems and methods of spectrophotometry for the determination of genome content, capsid content and full/empty ratios of adeno-associated virus particles
WO2019212921A1 (en) 2018-04-29 2019-11-07 Regenxbio Inc. Scalable clarification process for recombinant aav production
BR112020022858A2 (pt) 2018-05-11 2021-02-23 Massachusetts Eye And Ear Infirmary tropismo específico hepático de vírus adeno-associado
US11821009B2 (en) 2018-05-15 2023-11-21 Cornell University Genetic modification of the AAV capsid resulting in altered tropism and enhanced vector delivery
EP3808840A4 (de) * 2018-06-14 2022-03-23 Avixgen Inc. Pharmazeutische zusammensetzung enthaltend ein fusionsprotein aus einem zellpenetrierenden peptid und rpe65 zur behandlung von genitalamaurosis
CA3102817A1 (en) 2018-06-14 2019-12-19 Claire G. ZHANG Anion exchange chromatography for recombinant aav production
CN112703198A (zh) 2018-07-11 2021-04-23 布里格姆妇女医院 用于跨血脑屏障递送试剂的方法和组合物
CN112567027A (zh) 2018-08-10 2021-03-26 再生生物股份有限公司 用于重组aav生产的可扩展方法
EP3856913A4 (de) 2018-09-26 2022-10-26 California Institute Of Technology Adeno-assoziierte viruszusammensetzungen zur gezielten gentherapie
CA3115524A1 (en) 2018-10-15 2020-04-23 Win Den Cheung Methods for measuring the infectivity of replication defective viral vectors and viruses
US20220090129A1 (en) * 2018-12-05 2022-03-24 Abeona Therapeutics Inc. Recombinant adeno-associated viral vector for gene delivery
EP3893717A4 (de) * 2018-12-12 2022-09-14 Tesseract Health, Inc. Biometrische identifizierungsverfahren
EP3897679A1 (de) * 2018-12-21 2021-10-27 Wade Wei-De Chien Adeno-assoziierte viren und deren verwendungen zur innenohrtherapie
CN110437317B (zh) * 2019-01-30 2023-05-02 上海科技大学 具有变异衣壳蛋白的腺相关病毒及其用途
KR20210126096A (ko) 2019-02-15 2021-10-19 상가모 테라퓨틱스, 인코포레이티드 재조합 aav를 제조하기 위한 조성물 및 방법
EP3931335A2 (de) 2019-02-25 2022-01-05 Novartis AG Zusammensetzungen und verfahren zur behandlung von bietti-kristall-dystrophie
SG11202108044YA (en) 2019-02-25 2021-09-29 Novartis Ag Compositions and methods to treat bietti crystalline dystrophy
CN113557010A (zh) 2019-02-28 2021-10-26 瑞泽恩制药公司 用于递送治疗剂的腺相关病毒载体
CN111621502B (zh) * 2019-02-28 2023-05-02 武汉纽福斯生物科技有限公司 视网膜劈裂蛋白的编码序列、其表达载体构建及其应用
KR20210135267A (ko) 2019-03-04 2021-11-12 애드베룸 바이오테크놀로지스, 인코포레이티드 반대측 눈에 대한 aav 유전자 요법의 순차적 유리체내 투여
US20220143221A1 (en) 2019-04-03 2022-05-12 Regenxbio, Inc. Gene Therapy For Eye Pathologies
TW202102526A (zh) * 2019-04-04 2021-01-16 美商銳進科斯生物股份有限公司 重組腺相關病毒及其用途
PL3953483T3 (pl) 2019-04-11 2024-04-22 Regenxbio Inc. Sposoby chromatografii wykluczania do charakteryzowania kompozycji rekombinowanych wirusów towarzyszących adenowirusom
CA3136939A1 (en) 2019-04-19 2020-10-22 Regenxbio Inc. Adeno-associated virus vector formulations and methods
JP2022530006A (ja) 2019-04-24 2022-06-27 リジェネックスバイオ インコーポレイテッド 完全ヒト翻訳後修飾抗体による治療剤
WO2020222858A1 (en) * 2019-04-27 2020-11-05 Ocugen, Inc. Adeno-associated virus vector mediated gene therapy for ophthalmic diseases
US11737665B2 (en) 2019-06-21 2023-08-29 Tesseract Health, Inc. Multi-modal eye imaging with shared optical path
JP2022539156A (ja) * 2019-06-27 2022-09-07 ユニバーシティー オブ フロリダ リサーチ ファンデーション， インク． ヒアルロン酸を用いた眼組織へのａａｖ媒介形質導入の増強
EP3997225A1 (de) 2019-07-10 2022-05-18 INSERM (Institut National de la Santé et de la Recherche Médicale) Verfahren zur behandlung von epilepsie
JP2022540226A (ja) * 2019-07-11 2022-09-14 サントル ナショナル ドゥ ラ ルシェルシュ シアンティフィック 化学修飾型アデノ随伴ウイルス
US10653731B1 (en) 2019-07-15 2020-05-19 Vigene Biosciences Inc. Recombinantly-modified adeno-associated virus (rAAV) having improved packaging efficiency
US10801042B1 (en) 2019-07-15 2020-10-13 Vigene Biosciences, Inc. Use of ion concentrations to increase the packaging efficiency of recombinant adeno-associated virus
US10557149B1 (en) 2019-07-15 2020-02-11 Vigene Biosciences, Inc. Recombinantly-modified adeno-associated virus helper vectors and their use to improve the packaging efficiency of recombinantly-modified adeno-associated virus
WO2021021661A1 (en) 2019-07-26 2021-02-04 Regenxbio Inc. Engineered nucleic acid regulatory element and methods of uses thereof
RU2751592C2 (ru) 2019-08-22 2021-07-15 Общество С Ограниченной Ответственностью "Анабион" Выделенный модифицированный белок VP1 капсида аденоассоциированного вируса 5 серотипа (AAV5), капсид и вектор на его основе
BR112022003811A2 (pt) 2019-08-26 2022-11-16 Regenxbio Inc Tratamento de retinopatia diabética com fab anti vegf totalmente humano modificado pós tradução
WO2021050649A1 (en) 2019-09-11 2021-03-18 Adverum Biotechnologies, Inc. Methods of treating ocular neovascular diseases using aav2 variants encoding aflibercept
AU2020347276A1 (en) * 2019-09-12 2022-03-17 Massachusetts Institute Of Technology Engineered adeno-associated virus capsids
GB201913974D0 (en) 2019-09-27 2019-11-13 King S College London Vector
US20220347298A1 (en) 2019-10-04 2022-11-03 Ultragenyx Pharmaceutical Inc. Methods for improved therapeutic use of recombinant aav
CN114728049A (zh) 2019-10-07 2022-07-08 再生生物股份有限公司 腺相关病毒载体药物组合物和方法
CN113518824B (zh) * 2019-10-16 2024-02-23 上海药明康德新药开发有限公司 新的aav变体
BR112022008861A2 (pt) * 2019-11-08 2022-08-23 Regeneron Pharma Construto de ácido nucleico, vetor, nanopartícula lipídica, célula, composição para uso na expressão de retinosquisina, método para integrar uma sequência de codificação, e, método para modificar um gene rs1
WO2021099394A1 (en) 2019-11-19 2021-05-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Antisense oligonucleotides and their use for the treatment of cancer
US20230057380A1 (en) * 2019-11-26 2023-02-23 University Of Massachusetts Recombinant adeno-associated virus for delivery of kh902 (conbercept) and uses thereof
MX2022006427A (es) 2019-11-28 2022-09-07 Regenxbio Inc "construcciones para terapia génica con microdistrofina y uso de las mismas.
KR20220127847A (ko) 2020-01-10 2022-09-20 더 브리검 앤드 우먼즈 하스피털, 인크. 뇌암을 치료하기 위해 혈액-뇌 장벽을 통해 면역요법제를 전달하기 위한 방법 및 조성물
TW202140791A (zh) 2020-01-13 2021-11-01 美商霍蒙拉奇醫藥公司 治療苯酮尿症之方法
KR20220131522A (ko) 2020-01-22 2022-09-28 리젠엑스바이오 인크. 완전-인간 글리코실화된 인간 알파-l-이두로니다아제 (idua)를 사용한 점액다당류증 i의 치료
WO2021154956A1 (en) 2020-01-29 2021-08-05 Regenxbio Inc. Treatment of mucopolysaccharidosis iva
AU2021214174A1 (en) 2020-01-29 2022-08-18 Regenxbio Inc. Treatment of mucopolysaccharidosis II with recombinant human iduronate-2-sulfatase (IDS) produced by human neural or glial cells
WO2021163322A1 (en) 2020-02-14 2021-08-19 Ultragenyx Pharmaceutical Inc. Gene therapy for treating cdkl5 deficiency disorder
US20230235353A1 (en) 2020-03-19 2023-07-27 Ultragenyx Pharmaceutical Inc. Compositions and methods for reducing reverse packaging of cap and rep sequences in recombinant aav
EP4127189A1 (de) 2020-03-31 2023-02-08 Ultragenyx Pharmaceutical Inc. Gentherapie zur behandlung von propionischer azidämie
WO2021204407A1 (en) 2020-04-10 2021-10-14 Sorbonne Université G-protein-gated-k+ channel-mediated enhancements in light sensitivity in rod-cone dystrophy (rcd)
JP2023523211A (ja) 2020-04-24 2023-06-02 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル 眼科遺伝子治療後の、導入遺伝子産物を発現する形質転換細胞に対する免疫応答の誘導を防止するための方法
CN115515613A (zh) 2020-04-27 2022-12-23 4D分子治疗有限公司 密码子优化的gla基因及其用途
JP2023525810A (ja) 2020-05-13 2023-06-19 ボイジャー セラピューティクス インコーポレイテッド Ａａｖカプシドの指向性の再指示
US20240018524A1 (en) 2020-07-10 2024-01-18 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and compositions for treating epilepsy
EP4199972A2 (de) * 2020-08-21 2023-06-28 Capsida, Inc. Adeno-assoziierte viruszusammensetzungen mit bevorzugten expressionsniveaus
CN112226461B (zh) * 2020-08-21 2022-04-22 华侨大学 Cd4阳性细胞特异性基因传递载体及其应用
WO2022060915A1 (en) 2020-09-15 2022-03-24 Regenxbio Inc. Vectorized lanadelumab and administration thereof
EP4214242A1 (de) 2020-09-15 2023-07-26 RegenxBio Inc. Vektorisierte antikörper für antivirale therapie
CN113754727B (zh) * 2020-09-30 2022-07-12 广州派真生物技术有限公司 腺相关病毒突变体及其应用
EP4225381A1 (de) 2020-10-07 2023-08-16 RegenxBio Inc. Formulierungen zur suprachoroidalen verabreichung wie formulierungen mit aggregatbildung
MX2023004005A (es) 2020-10-07 2023-06-06 Regenxbio Inc Formulaciones para administración supracoroidea tales como formulaciones de gel.
JP2023545722A (ja) 2020-10-07 2023-10-31 リジェネックスバイオ インコーポレイテッド 遺伝子治療剤の眼送達のためのアデノ随伴ウイルス
TW202228648A (zh) 2020-10-07 2022-08-01 美商銳進科斯生物股份有限公司 諸如高黏度調配物之用於脈絡膜上投與之調配物
WO2022076750A2 (en) 2020-10-07 2022-04-14 Regenxbio Inc. Recombinant adeno-associated viruses for cns or muscle delivery
WO2022094157A1 (en) 2020-10-28 2022-05-05 Regenxbio Inc. Vectorized anti-cgrp and anti-cgrpr antibodies and administration thereof
CA3195967A1 (en) 2020-10-28 2022-05-05 Xu Wang Vectorized anti-tnf-? antibodies for ocular indications
EP4236974A2 (de) 2020-10-29 2023-09-06 RegenxBio Inc. Vektorisierte faktor xii-antikörper und verabreichung davon
IL302282A (en) 2020-10-29 2023-06-01 Regenxbio Inc Vectored TNF-alpha antagonists for ocular indications
WO2022120022A1 (en) 2020-12-02 2022-06-09 Regeneron Pharmaceuticals, Inc. Crispr sam biosensor cell lines and methods of use thereof
US20240067678A1 (en) * 2020-12-16 2024-02-29 Children's Medical Research Institute Adeno-associated virus capsids and vectors
JP2023554389A (ja) 2020-12-16 2023-12-27 リジェネクスバイオ インコーポレイテッド 組換えアデノ随伴ウイルスウイルス粒子を生成する方法
TW202241943A (zh) 2020-12-29 2022-11-01 美商銳進科斯生物股份有限公司 Tau特異性抗體基因療法組合物、方法及其用途
KR20230133314A (ko) 2021-01-21 2023-09-19 리젠엑스바이오 인크. 재조합 폴리펩티드 및 바이러스의 개선된 생산
MX2023009118A (es) 2021-02-10 2023-10-18 Regenxbio Inc Tratamiento de la mucopolisacaridosis ii con iduronato-2-sulfatasa (ids) humana recombinante.
CN113121651B (zh) * 2021-04-19 2023-11-17 信念医药科技(上海)有限公司 低中和抗体腺相关病毒衣壳蛋白
CN113121652B (zh) * 2021-04-19 2022-10-11 上海信致医药科技有限公司 视网膜和肌肉高亲和性腺相关病毒衣壳蛋白及相关应用
WO2022223644A2 (en) 2021-04-20 2022-10-27 INSERM (Institut National de la Santé et de la Recherche Médicale) Compositions and methods for treating retinal degenerative disorders
JP2024517143A (ja) 2021-04-26 2024-04-19 リジェネックスバイオ インコーポレイテッド ジストロフィン異常症の治療のためのマイクロジストロフィン遺伝子療法投与
US20240197915A1 (en) 2021-04-27 2024-06-20 Adverum Biotechnologies, Inc. Methods of treating ocular diseases using aav2 variants encoding aflibercept
EP4334454A2 (de) 2021-05-04 2024-03-13 RegenxBio Inc. Neue aav-vektoren sowie verfahren und verwendungen davon
EP4337267A1 (de) 2021-05-11 2024-03-20 RegenxBio Inc. Behandlung von duchenne-muskeldystrophie und kombinationen davon
EP4347848A1 (de) 2021-05-26 2024-04-10 Sparingvision Durch g-protein-gated-k+-kanal vermittelte verbesserungen der lichtempfindlichkeit bei der rod-cone-dystrophie (rcd)
TW202313096A (zh) 2021-05-28 2023-04-01 大陸商江蘇恆瑞醫藥股份有限公司 衣殼變異的重組腺相關病毒及其應用
AU2022283898A1 (en) * 2021-06-03 2024-01-04 Dyno Therapeutics, Inc. Capsid variants and methods of using the same
WO2023280926A1 (en) 2021-07-07 2023-01-12 INSERM (Institut National de la Santé et de la Recherche Médicale) Synergistic combination of rdcfv2 and rdcvf2l for the treatment of tauopathies
TW202317768A (zh) 2021-07-08 2023-05-01 美商特納亞治療股份有限公司 用於基因療法的優化表現卡匣
CA3226452A1 (en) 2021-07-19 2023-01-26 New York University Auf1 combination therapies for treatment of muscle degenerative disease
WO2023019168A1 (en) 2021-08-11 2023-02-16 Ultragenyx Pharmaceutical Inc. Compositions and methods for treating a muscular dystrophy
AR126839A1 (es) 2021-08-20 2023-11-22 Llc «Anabion» Proteína de la cápside vp1 modificada aislada del virus adeno-asociado de serotipo 9 (aav9), cápside y vector basado en esta
AU2022328531A1 (en) 2021-08-20 2024-03-14 Joint Stock Company "Biocad" Method of obtaining a modified adeno-associated virus capsid
AR126840A1 (es) 2021-08-20 2023-11-22 Llc «Anabion» Proteína de la cápside vp1 modificada aislada del virus adeno-asociado de serotipo 5 (aav5), cápside y vector basado en esta
WO2023060113A1 (en) 2021-10-05 2023-04-13 Regenxbio Inc. Compositions and methods for recombinant aav production
CN118202060A (zh) 2021-10-05 2024-06-14 再生生物股份有限公司 用于重组aav生产的组合物和方法
WO2023060269A1 (en) 2021-10-07 2023-04-13 Regenxbio Inc. Recombinant adeno-associated viruses for targeted delivery
WO2023060272A2 (en) 2021-10-07 2023-04-13 Regenxbio Inc. Recombinant adeno-associated viruses for cns tropic delivery
EP4163296A1 (de) 2021-10-11 2023-04-12 Sparingvision Durch g-protein-gated-k+-kanal vermittelte verbesserungen der lichtempfindlichkeit bei patienten mit der zapfen-stäbchen-dystrophie
WO2023077092A1 (en) 2021-10-28 2023-05-04 Regenxbio Inc. Engineered nucleic acid regulatory elements and methods and uses thereof
WO2023108507A1 (en) * 2021-12-15 2023-06-22 National Institute Of Biological Sciences, Beijing Recombinant aav vectors and use thereof
WO2023150566A1 (en) 2022-02-02 2023-08-10 Adverum Biotechnologies, Inc. Methods of treating ocular neovascular diseases using aav2 variants encoding aflibercept
WO2023158990A1 (en) 2022-02-16 2023-08-24 Adverum Biotechnologies, Inc. Method of reducing cst fluctuation in neovascular amd by a recombinant adeno-associated virus
WO2023155828A1 (en) 2022-02-17 2023-08-24 Skyline Therapeutics (Shanghai) Co., Ltd. Recombinant adeno-associated virus with modified aav capsid polypeptides
TW202342740A (zh) 2022-03-07 2023-11-01 美商奧崔基尼克斯製藥公司 改良的批量aav生產系統和方法
WO2023178053A1 (en) 2022-03-13 2023-09-21 Regenxbio Inc. Modified muscle-specific promoters
WO2023183623A1 (en) 2022-03-25 2023-09-28 Regenxbio Inc. Dominant-negative tumor necrosis factor alpha adeno-associated virus gene therapy
WO2023196873A1 (en) 2022-04-06 2023-10-12 Regenxbio Inc. Pharmaceutical composition comprising a recombinant adeno-associated virus vector with an expression cassette encoding a transgene forsuprachoidal administration
TW202404651A (zh) 2022-04-06 2024-02-01 美商銳進科斯生物股份有限公司 用於脈絡膜上投與之調配物諸如形成聚集體之調配物
TW202345913A (zh) 2022-04-06 2023-12-01 美商銳進科斯生物股份有限公司 用於脈絡膜上投與之調配物諸如凝膠調配物
TW202404993A (zh) 2022-04-11 2024-02-01 美商特納亞治療股份有限公司 具經工程化蛋白殼之腺相關病毒
WO2023201308A1 (en) 2022-04-14 2023-10-19 Regenxbio Inc. Gene therapy for treating an ocular disease
WO2023201277A1 (en) 2022-04-14 2023-10-19 Regenxbio Inc. Recombinant adeno-associated viruses for cns tropic delivery
CN116970648A (zh) * 2022-04-24 2023-10-31 上海朗昇生物科技有限公司 新型aav衣壳改造株及其用途
WO2023215807A1 (en) 2022-05-03 2023-11-09 Regenxbio Inc. VECTORIZED ANTI-TNF-α INHIBITORS FOR OCULAR INDICATIONS
TW202400803A (zh) 2022-05-03 2024-01-01 美商銳進科斯生物股份有限公司 載體化抗補體抗體與補體劑及其投與
WO2023240062A1 (en) 2022-06-07 2023-12-14 Adverum Biotechnologies, Inc. Melanopsin variants for vision restoration
WO2023239627A2 (en) 2022-06-08 2023-12-14 Regenxbio Inc. Methods for recombinant aav production
WO2024017990A1 (en) 2022-07-21 2024-01-25 Institut National de la Santé et de la Recherche Médicale Methods and compositions for treating chronic pain disorders
WO2024033837A1 (en) 2022-08-11 2024-02-15 Institute Of Molecular And Clinical Ophthalmology Basel (Iob) Human cone photoreceptor optogenetic constructs
WO2024044725A2 (en) 2022-08-24 2024-02-29 Regenxbio Inc. Recombinant adeno-associated viruses and uses thereof
WO2024073669A1 (en) 2022-09-30 2024-04-04 Regenxbio Inc. Treatment of ocular diseases with recombinant viral vectors encoding anti-vegf fab
WO2024081746A2 (en) 2022-10-11 2024-04-18 Regenxbio Inc. Engineered nucleic acid regulatory elements and methods and uses thereof
WO2024084075A1 (en) 2022-10-20 2024-04-25 Sparingvision Compositions and methods for treating retinal degenerative disorders
EP4357359A1 (de) 2022-10-20 2024-04-24 Sparingvision Durch g-protein-gated-k+-kanal vermittelte verbesserungen der lichtempfindlichkeit bei der rod-cone-dystrophie (rcd)
WO2024107670A1 (en) 2022-11-16 2024-05-23 Regeneron Pharmaceuticals, Inc. Chimeric proteins comprising membrane bound il-12 with protease cleavable linkers

Citations (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5639736A (en) 1993-07-27 1997-06-17 Hybridon, Inc. Human VEGF-specific oligonucleotides
WO1998011244A2 (en) 1996-09-11 1998-03-19 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Aav4 vector and uses thereof
WO1999061601A2 (en) 1998-05-28 1999-12-02 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Aav5 vector and uses thereof
WO2000028061A2 (en) 1998-11-05 2000-05-18 The Trustees Of The University Of Pennsylvania Adeno-associated virus serotype 1 nucleic acid sequences, vectors and host cells containing same
US6156303A (en) 1997-06-11 2000-12-05 University Of Washington Adeno-associated virus (AAV) isolates and AAV vectors derived therefrom
US20010008642A1 (en) 2000-01-14 2001-07-19 Ulrich Meyer Extruder die head
US6399586B1 (en) 1993-07-27 2002-06-04 Hybridon, Inc. Modified VEGF oligonucleotides
US20050053922A1 (en) 2003-06-30 2005-03-10 Schaffer David V. Mutant adeno-associated virus virions and methods of use thereof
US7271002B2 (en) 2001-11-09 2007-09-18 United States Of America, Represented By The Secretary, Department Of Health And Human Services Production of adeno-associated virus in insect cells
US20070261127A1 (en) 2005-07-22 2007-11-08 Boyden Edward S Light-activated cation channel and uses thereof
US20080188437A1 (en) 2002-07-24 2008-08-07 The Trustees Of The University Of Pennsylvania Compositions and Methods for siRNA Inhibition of Angiogenesis
US20090202490A1 (en) 2003-06-30 2009-08-13 Schaffer David V Mutant adeno-associated virus virions and methods of use thereof
US20100015095A1 (en) 2006-05-04 2010-01-21 Wayne State University Restoration of visual responses by in vivo delivery of rhodopsin nucleic acids
WO2010093784A2 (en) * 2009-02-11 2010-08-19 The University Of North Carolina At Chapel Hill Modified virus vectors and methods of making and using the same
US7846730B2 (en) 2001-07-17 2010-12-07 Isis Pharmaceuticals, Inc. Antisense modulation of BCL2-associated X protein expression
US7919473B2 (en) 2004-03-12 2011-04-05 Alnylam Pharmaceuticals, Inc. IRNA agents targeting VEGF
US20110143400A1 (en) 2006-09-08 2011-06-16 Opko Ophthalmics, Llc Sirna and methods of manufacture
US20110301073A1 (en) 2010-05-17 2011-12-08 Sangamo Biosciences, Inc. Novel DNA-binding proteins and uses thereof

Family Cites Families (83)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9206016D0 (en) 1992-03-19 1992-04-29 Sandoz Ltd Improvements in or relating to organic compounds
US6440425B1 (en) 1995-05-01 2002-08-27 Aventis Pasteur Limited High molecular weight major outer membrane protein of moraxella
US6001650A (en) * 1995-08-03 1999-12-14 Avigen, Inc. High-efficiency wild-type-free AAV helper functions
US6096548A (en) 1996-03-25 2000-08-01 Maxygen, Inc. Method for directing evolution of a virus
CA2251738A1 (en) 1996-04-16 1997-10-23 Immusol Incorporated Targeted viral vectors
US6710036B2 (en) 1997-07-25 2004-03-23 Avigen, Inc. Induction of immune response to antigens expressed by recombinant adeno-associated virus
WO1999014354A1 (en) 1997-09-19 1999-03-25 The Trustees Of The University Of The Pennsylvania Methods and vector constructs useful for production of recombinant aav
EP1030861A4 (de) 1997-10-31 2001-09-05 Maxygen Inc Modifizierung des virustropismius und des wirtspektrums durch virale genomdurchmischung
US6410300B1 (en) 1998-01-12 2002-06-25 The University Of North Carolina At Chapel Hill Methods and formulations for mediating adeno-associated virus (AAV) attachment and infection and methods for purifying AAV
US6551795B1 (en) * 1998-02-18 2003-04-22 Genome Therapeutics Corporation Nucleic acid and amino acid sequences relating to pseudomonas aeruginosa for diagnostics and therapeutics
DE19827457C1 (de) 1998-06-19 2000-03-02 Medigene Ag Strukturprotein von AAV, seine Herstellung und Verwendung
EP1109925B1 (de) * 1998-09-11 2005-05-04 The Regents of The University of California Rekombinante adenovirus für gewebespezifische genexpression ins herz
JP2002538770A (ja) 1998-11-10 2002-11-19 ユニバーシティ オブ ノース カロライナ アット チャペル ヒル ウイルスベクターとその製造及び投与の方法
US6943153B1 (en) 1999-03-15 2005-09-13 The Regents Of The University Of California Use of recombinant gene delivery vectors for treating or preventing diseases of the eye
US6498244B1 (en) 1999-05-28 2002-12-24 Cell Genesys, Inc. Adeno-associated virus capsid immunologic determinants
US7314912B1 (en) 1999-06-21 2008-01-01 Medigene Aktiengesellschaft AAv scleroprotein, production and use thereof
DE19933288A1 (de) 1999-07-15 2001-01-18 Medigene Ag Strukturprotein von Adeno-assoziiertem Virus mit veränderter Antigenität, seine Herstellung und Verwendung
DE19933719A1 (de) 1999-07-19 2001-01-25 Medigene Ag Strukturprotein in Adeno-assoziiertem Virus mit veränderten chromatographischen Eigenschaften, seine Herstellung und Verwendung
US8232081B2 (en) 1999-09-21 2012-07-31 Basf Se Methods and microorganisms for production of panto-compounds
WO2002081628A2 (en) 2001-04-05 2002-10-17 Ribozyme Pharmaceuticals, Incorporated Modulation of gene expression associated with inflammation proliferation and neurite outgrowth, using nucleic acid based technologies
US6855314B1 (en) 2000-03-22 2005-02-15 The United States Of America As Represented By The Department Of Health And Human Services AAV5 vector for transducing brain cells and lung cells
IL154987A0 (en) * 2000-09-22 2003-10-31 Virxsys Conditionally replicating viral vectors and their use
US7749492B2 (en) * 2001-01-05 2010-07-06 Nationwide Children's Hospital, Inc. AAV vectors and methods
US6962815B2 (en) * 2001-01-05 2005-11-08 Children's Hopital Inc. AAV2 vectors and methods
US20030129203A1 (en) * 2001-08-27 2003-07-10 Nautilus Biotech S.A. Mutant recombinant adeno-associated viruses
US7647184B2 (en) 2001-08-27 2010-01-12 Hanall Pharmaceuticals, Co. Ltd High throughput directed evolution by rational mutagenesis
EP2338900B1 (de) 2001-11-13 2014-01-01 The Trustees of The University of Pennsylvania Adenoassoziierten Virus (AAV) Cy.5 Sequenzen, diese enthaltende Vektoren und deren Verwendungen
ES2467156T3 (es) 2001-12-21 2014-06-12 Medigene Ag Una biblioteca de genes estructurales modificados o de partículas modificadas de cápside útiles para la identificación de clones virales con tropismo celular deseado
DE10216005A1 (de) 2002-04-11 2003-10-30 Max Planck Gesellschaft Verwendung von biologischen Photorezeptoren als direkt lichtgesteuerte Ionenkanäle
WO2003089612A2 (en) 2002-04-17 2003-10-30 University Of Florida Research Foundation, Inc. IMPROVED rAAV VECTORS
AU2003237159A1 (en) 2002-04-30 2003-11-17 University Of Florida Treatment for phenylketonuria
US7858367B2 (en) 2002-04-30 2010-12-28 Duke University Viral vectors and methods for producing and using the same
US7254489B2 (en) 2002-05-31 2007-08-07 Microsoft Corporation Systems, methods and apparatus for reconstructing phylogentic trees
WO2004020600A2 (en) 2002-08-28 2004-03-11 University Of Florida Modified aav
US20060223756A1 (en) * 2002-12-18 2006-10-05 Gene Liau Endothelial cell specifically binding peptides
WO2004083441A2 (en) 2003-03-19 2004-09-30 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts Random peptide library displayed on aav vectors
WO2004108922A2 (en) 2003-04-25 2004-12-16 The Trustees Of The University Of Pennsylvania Use of aav comprising a capsid protein from aav7 or aav8 for the delivery of genes encoding apoprotein a or e genes to the liver
US7611868B2 (en) * 2003-05-14 2009-11-03 Instituto Di Ricerche Di Biologia Molecolare P. Angeletti S.P.A. Recombinant modified adenovirus fiber protein
EP1486567A1 (de) 2003-06-11 2004-12-15 Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts Verbesserter Adeno-assoziierter Virus (AAV)-Vektor für Gentherapie
PT2657247T (pt) 2003-06-19 2017-06-29 Genzyme Corp Viriões aav com imunorreatividade diminuída e seus usos
US7368428B2 (en) 2003-06-23 2008-05-06 A&G Pharmaceutical, Inc. Compositions and methods for restoring sensitivity of tumor cells to antitumor therapy and inducing apoptosis
CA2539947A1 (en) 2003-09-24 2005-04-07 The Board Of Trustees Of The Leland Stanford Junior University Igf-1 instructs multipotent adult cns neural stem cells to an oligodendroglial lineage
CN102199626B (zh) 2003-09-30 2015-06-24 宾夕法尼亚大学托管会 腺伴随病毒(aav)进化支、序列、含有这些序列的载体及它们的应用
KR100550967B1 (ko) 2003-12-03 2006-02-13 주식회사 엘지생활건강 유중수형 화장료 조성물
US7427396B2 (en) 2004-06-03 2008-09-23 Genzyme Corporation AAV vectors for gene delivery to the lung
AU2005316476A1 (en) 2004-12-15 2006-06-22 University Of Florida Research Foundation, Inc. Chimeric vectors
US20060211650A1 (en) 2004-12-16 2006-09-21 Forest Laboratories, Inc. Reducing carbohydrate derivatives of adamantane amines, and synthesis and methods of use thereof
WO2006101634A1 (en) 2005-02-17 2006-09-28 The Regents Of The University Of California Müller cell specific gene therapy
CN101203613B (zh) 2005-04-07 2012-12-12 宾夕法尼亚大学托管会 增强腺相关病毒载体功能的方法
CN1834255A (zh) * 2005-12-23 2006-09-20 上海交通大学附属第一人民医院 加快并提高腺相关病毒介导的基因在视网膜细胞内表达的方法
WO2007089632A2 (en) 2006-01-27 2007-08-09 The University Of North Carolina At Chapel Hill Heparin and heparan sulfate binding chimeric vectors
WO2007120542A2 (en) 2006-03-30 2007-10-25 The Board Of Trustees Of The Leland Stanford Junior University Aav capsid library and aav capsid proteins
CN101506369B (zh) * 2006-06-21 2014-02-12 尤尼克尔生物制药股份有限公司 具有经修饰的用于在昆虫细胞中产生aav的aav-rep78翻译起始密码子的载体
CN1966082B (zh) * 2006-11-03 2010-06-30 许瑞安 一种防治结直肠癌的基因药物及其制备方法和用途
US9725485B2 (en) 2012-05-15 2017-08-08 University Of Florida Research Foundation, Inc. AAV vectors with high transduction efficiency and uses thereof for gene therapy
PT2147096E (pt) 2007-04-13 2015-07-21 Catalyst Biosciences Inc Polipéptidos do factor vii modificados e suas utilizações
EP1985708B1 (de) * 2007-04-27 2015-04-15 Universität Rostock Selektives Targeting von Viren in neurale Vorläuferzellen
US8632764B2 (en) * 2008-04-30 2014-01-21 University Of North Carolina At Chapel Hill Directed evolution and in vivo panning of virus vectors
AU2009274482A1 (en) 2008-05-20 2010-01-28 Eos Neuroscience, Inc. Vectors for delivery of light-sensitive proteins and methods of use
WO2009154452A1 (en) 2008-06-17 2009-12-23 Amsterdam Molecular Therapeutics B.V. Parvoviral capsid with incorporated Gly-Ala repeat region
CN101532024A (zh) * 2009-04-30 2009-09-16 许瑞安 一种用于基因治疗的新型细胞特异性内含microRNA结合序列的基因HAAVmir
WO2010138263A2 (en) 2009-05-28 2010-12-02 University Of Massachusetts Novel aav 's and uses thereof
WO2011117258A2 (en) 2010-03-22 2011-09-29 Association Institut De Myologie Methods of increasing efficiency of vector penetration of target tissue
US8663624B2 (en) 2010-10-06 2014-03-04 The Regents Of The University Of California Adeno-associated virus virions with variant capsid and methods of use thereof
WO2012109570A1 (en) * 2011-02-10 2012-08-16 The University Of North Carolina At Chapel Hill Viral vectors with modified transduction profiles and methods of making and using the same
KR20220056884A (ko) * 2011-04-22 2022-05-06 더 리젠츠 오브 더 유니버시티 오브 캘리포니아 변이체 캡시드를 지니는 아데노-관련 바이러스 및 이의 사용 방법
US9169299B2 (en) 2011-08-24 2015-10-27 The Board Of Trustees Of The Leleand Stanford Junior University AAV capsid proteins for nucleic acid transfer
JP6385920B2 (ja) 2012-05-09 2018-09-05 オレゴン ヘルス アンド サイエンス ユニバーシティー アデノ随伴ウイルスプラスミド及びベクター
TWI702955B (zh) 2012-05-15 2020-09-01 澳大利亞商艾佛蘭屈澳洲私營有限公司 使用腺相關病毒（ａａｖ）ｓｆｌｔ-１治療老年性黃斑部退化（ａｍｄ）
CN104937100B (zh) 2012-12-25 2020-04-03 宝生物工程株式会社 Aav 变体
US9567376B2 (en) 2013-02-08 2017-02-14 The Trustees Of The University Of Pennsylvania Enhanced AAV-mediated gene transfer for retinal therapies
US11685935B2 (en) 2013-05-29 2023-06-27 Cellectis Compact scaffold of Cas9 in the type II CRISPR system
ES2897508T3 (es) 2013-05-31 2022-03-01 Univ California Variantes de virus adenoasociados y métodos de uso de las mismas
CA2930549A1 (en) 2013-09-26 2015-04-02 University Of Florida Research Foundation, Inc. Synthetic combinatorial aav capsid library for targeted gene therapy
EP3060575B1 (de) 2013-10-11 2018-12-05 Massachusetts Eye & Ear Infirmary Verfahren zur vorhersage anzestraler virussequenzen und verwendungen davon
EP3800191A1 (de) 2014-03-17 2021-04-07 Adverum Biotechnologies, Inc. Zusammensetzungen und verfahren zur verbesserten genexpression in zapfenzellen
AU2015252797C1 (en) 2014-05-02 2021-04-22 Genzyme Corporation AAV vectors for retinal and CNS gene therapy
EP3155116A4 (de) 2014-06-10 2017-12-27 Massachusetts Institute Of Technology Verfahren zur genbearbeitung
AU2016226289B2 (en) 2015-03-02 2021-04-29 Adverum Biotechnologies, Inc. Compositions and methods for intravitreal delivery of polynucleotides to retinal cones
HUE051491T2 (hu) 2015-03-06 2021-03-01 Massachusetts Eye & Ear Infirmary Gén-augmentációs terápiák a PRPF31 gén mutációi által okozott öröklött retina degenerációra
WO2017023724A1 (en) 2015-07-31 2017-02-09 Voyager Therapeutics, Inc. Compositions and methods for the treatment of aadc deficiency
CN109476707B (zh) 2016-05-13 2022-12-02 4D分子治疗有限公司 腺相关病毒变体衣壳和其使用方法
US11680249B2 (en) 2017-08-28 2023-06-20 The Regents Of The University Of California Adeno-associated virus capsid variants and methods of use thereof

Patent Citations (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6399586B1 (en) 1993-07-27 2002-06-04 Hybridon, Inc. Modified VEGF oligonucleotides
US5639872A (en) 1993-07-27 1997-06-17 Hybridon, Inc. Human VEGF-specific oligonucleotides
US5661135A (en) 1993-07-27 1997-08-26 Hybridon, Inc. Human VEGF-specific oligonucleotides
US5639736A (en) 1993-07-27 1997-06-17 Hybridon, Inc. Human VEGF-specific oligonucleotides
US6649596B1 (en) 1995-12-08 2003-11-18 Hybridon, Inc. Modified VEGF Oligonucleotides for Inhibition of tumor growth
WO1998011244A2 (en) 1996-09-11 1998-03-19 The Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Aav4 vector and uses thereof
US6156303A (en) 1997-06-11 2000-12-05 University Of Washington Adeno-associated virus (AAV) isolates and AAV vectors derived therefrom
WO1999061601A2 (en) 1998-05-28 1999-12-02 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Aav5 vector and uses thereof
WO2000028061A2 (en) 1998-11-05 2000-05-18 The Trustees Of The University Of Pennsylvania Adeno-associated virus serotype 1 nucleic acid sequences, vectors and host cells containing same
US20010008642A1 (en) 2000-01-14 2001-07-19 Ulrich Meyer Extruder die head
US7846730B2 (en) 2001-07-17 2010-12-07 Isis Pharmaceuticals, Inc. Antisense modulation of BCL2-associated X protein expression
US7271002B2 (en) 2001-11-09 2007-09-18 United States Of America, Represented By The Secretary, Department Of Health And Human Services Production of adeno-associated virus in insect cells
US20080188437A1 (en) 2002-07-24 2008-08-07 The Trustees Of The University Of Pennsylvania Compositions and Methods for siRNA Inhibition of Angiogenesis
US20050053922A1 (en) 2003-06-30 2005-03-10 Schaffer David V. Mutant adeno-associated virus virions and methods of use thereof
US20090202490A1 (en) 2003-06-30 2009-08-13 Schaffer David V Mutant adeno-associated virus virions and methods of use thereof
US7919473B2 (en) 2004-03-12 2011-04-05 Alnylam Pharmaceuticals, Inc. IRNA agents targeting VEGF
US7947659B2 (en) 2004-03-12 2011-05-24 Alnylam Pharmaceuticals, Inc. iRNA agents targeting VEGF
US20070261127A1 (en) 2005-07-22 2007-11-08 Boyden Edward S Light-activated cation channel and uses thereof
US20100015095A1 (en) 2006-05-04 2010-01-21 Wayne State University Restoration of visual responses by in vivo delivery of rhodopsin nucleic acids
US20110143400A1 (en) 2006-09-08 2011-06-16 Opko Ophthalmics, Llc Sirna and methods of manufacture
WO2010093784A2 (en) * 2009-02-11 2010-08-19 The University Of North Carolina At Chapel Hill Modified virus vectors and methods of making and using the same
US20110301073A1 (en) 2010-05-17 2011-12-08 Sangamo Biosciences, Inc. Novel DNA-binding proteins and uses thereof

Non-Patent Citations (63)

* Cited by examiner, † Cited by third party
Title
"GenBank", Database accession no. NP 001289
"Macromolecule Sequencing and Synthesis, Selected Methods and Applications", 1988, ALAN R. LISS, INC., article "Current Methods in Sequence Comparison and Analysis", pages: 127 - 149
"Methods in Enzymology", vol. 266, 1996, ACADEMIC PRESS, INC., article "Computer Methods for Macromolecular Sequence Analysis"
"Pharmaceutical Dosage Forms and Drug Delivery Systems", 1999, LIPPINCOTT, WILLIAMS, & WILKINS
AKIYAMA ET AL., J. CELL PHYSIOL., vol. 207, 2006, pages 407
ALLOCCA ET AL., J. VIROL., vol. 81, 2007, pages 11372
BANTEL-SCHAAL ET AL., J. VIROLOGY, vol. 73, 1999, pages 3994
BOOIJ ET AL., OPHTHALMOLOGY, vol. 118, 2011, pages 160 - 167
BOUCAS ET AL., J. GENE MED., vol. 11, 2009, pages 1103
CHIORINI ET AL., J. VIROLOGY, vol. 71, 1998, pages 6823
DAIGER SPBOWNE SJSULLIVAN LS: "Perspective on genes and mutations causing retinitis pigmentosa", ARCH OPHTHALMOL, vol. 125, 2007, pages 151 - 158
DALKARA DKOLSTAD KDCAPORALE NVISEL MKLIMCZAK RR ET AL.: "Inner Limiting Membrane Barriers to AAV Mediated Retinal Transduction from the Vitreous", MOL THER., 2009
DEN HOLLANDER AIROEPMAN RKOENEKOOP RKCREMERS FP: "Leber congenital amaurosis: genes, proteins and disease mechanisms", PROG RETIN EYE RES, vol. 27, 2008, pages 391 - 419, XP023611414, DOI: 10.1016/j.preteyeres.2008.05.003
DEN HOLLANDER ET AL., NAT. GENET., vol. 23, 1999, pages 217
DIESTER ET AL., NAT. NEUROSCI., vol. 14, 2011, pages 387
GAO ET AL., PROC. NAT. ACAD. SCI. USA, vol. 99, 2002, pages 11854
GRAINGER ET AL., MOL. THER., vol. 12, 2005, pages S337
GRUTER OKOSTIC CCRIPPA SVPEREZ MTZOGRAFOS L ET AL.: "Lentiviral vector-mediated gene transfer in adult mouse photoreceptors is impaired by the presence of a physical barrier", GENE THER, vol. 12, 2005, pages 942 - 947
HILDA PETRS-SILVA ET AL: "High-efficiency Transduction of the Mouse Retina by Tyrosine-mutant AAV Serotype Vectors", MOLECULAR THERAPY, vol. 17, no. 3, March 2009 (2009-03-01), pages 463 - 471, XP055076954, ISSN: 1525-0016, DOI: 10.1038/mt.2008.269 *
J. MOL. BIOL., vol. 48, 1970, pages 443 - 453
KAISER ET AL., AM. J. OPHTHALMOL., vol. 150, 2010, pages 33
KOHL ET AL., EUR J HUM GENET., vol. 13, no. 3, 2005, pages 302
KOU ET AL., BIOCHEM., vol. 44, 2005, pages 15064
LEE ET AL., PROC. NATL. ACAD. SCI. USA, vol. 102, 2005, pages 18902
LI ET AL., NATURE, vol. 475, 2011, pages 217
MAGUIRE AMSIMONELLI FPIERCE EAPUGH EN, JR.MINGOZZI F ET AL.: "Safety and efficacy of gene transfer for Leber's congenital amaurosis", N ENGL J MED, vol. 358, 2008, pages 2240 - 2248, XP055027817, DOI: 10.1056/NEJMoa0802315
MANCUSO KHAUSWIRTH WWLI QCONNOR TBKUCHENBECKER JA ET AL.: "Gene therapy for red-green colour blindness in adult primates", NATURE, vol. 461, no. 7265, 2009, pages 784 - 787, XP002615068, DOI: 10.1038/NATURE08401
MCGEE SANFTNER LHABEL HHAUSWIRTH WWFLANNERY JG: "Glial cell line derived neurotrophic factor delays photoreceptor degeneration in a transgenic rat model of retinitis pigmentosa", MOL THER, vol. 4, 2001, pages 622 - 629, XP002953220, DOI: 10.1006/mthe.2001.0498
METH. MOL. BIOL., vol. 70, 1997, pages 173 - 187
MORIMURA ET AL., PROC. NATL. ACAD. SCI. USA, vol. 95, 1998, pages 3088
MORIS ET AL., VIROLOGY, vol. 33, 2004, pages 375 - 383
MULLER OJKAUL FWEITZMAN MDPASQUALINI RARAP W ET AL.: "Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors", NAT BIOTECHNOL, vol. 21, 2003, pages 1040 - 1046, XP002596751, DOI: 10.1038/NBT856
MURAMATSU ET AL., VIROLOGY, vol. 221, 1996, pages 208
NAKAZAWA T. ET AL.: "Attenuated glial reactions and photoreceptor degeneration after retinal detachment in mice deficient in glial fibrillary acidic protein and vimentin", INVEST OPHTHAMOL VIS SCI, vol. 48, 2007, pages 2760 - 8
NAKAZAWA T. ET AL.: "Characterization of cytokine responses to retinal detachment in rats", MOL VIS, vol. 12, 2006, pages 867 - 78
NG ET AL., NAT. REV. DRUG DISCOVERY, vol. 5, 2006, pages 123
NICOUD ET AL., J. GENE MED., vol. 9, 2007, pages 1015
NIHUI, OPHTHALMOLOGICA, vol. 223, 2009, pages 401
PADRON ET AL., J. VIROL., vol. 79, 2005, pages 5047
PECHAN ET AL., GENE THER., vol. 16, 2009, pages 10
PERABO LBUNING HKOFLER DMRIED MUGIROD A ET AL.: "In vitro selection of viral vectors with modified tropism: the adeno-associated virus display", MOL THER, vol. 8, 2003, pages 151 - 157, XP003000206, DOI: 10.1016/S1525-0016(03)00123-0
PETRS-SILVA HDINCULESCU ALI QMIN SHCHIODO V ET AL.: "High-efficiency transduction of the mouse retina by tyrosine-mutant AAV serotype vectors", MOL THER, vol. 17, 2009, pages 463 - 471, XP055076957, DOI: 10.1038/mt.2008.269
REICH ET AL., MOL. VIS., vol. 9, 2003, pages 210
REME CEGRIMM CHAFEZI FWENZEL AWILLIAMS TP: "Apoptosis in the Retina: The Silent Death of Vision", NEWS PHYSIOL SCI, vol. 15, 2000, pages 120 - 124
ROLLING F: "Recombinant AAV-mediated gene transfer to the retina: gene therapy perspectives", GENE THER, vol. 11, no. 1, 2004, pages S26 - 32, XP002528464, DOI: 10.1038/SJ.GT.3302366
SAMBROOK ET AL.: "Molecular Cloning, A Laboratory Manual", 1989, COLD SPRING HARBOR LABORATORY PRESS
SHADE ET AL., J. VIROL., vol. 58, 1986, pages 921
SHEN ET AL., GENE THER., vol. 13, 2006, pages 225
SHEN ET AL., MOL. THER., vol. 15, 2007, pages 1955
SMITHWATERMAN, ADVANCES IN APPLIED MATHEMATICS, vol. 2, 1981, pages 482 - 489
SRIVISTAVA ET AL., J. VIROLOGY, vol. 45, 1983, pages 555
T K PARK ET AL: "Intravitreal delivery of AAV8 retinoschisin results in cell type-specific gene expression and retinal rescue in the Rs1-KO mouse", GENE THERAPY, vol. 16, no. 7, July 2009 (2009-07-01), pages 916 - 926, XP055113217, ISSN: 0969-7128, DOI: 10.1038/gt.2009.61 *
TRAVIS ET AL., GENOMICS, vol. 10, 1991, pages 733
VAN BOKHOVEN ET AL., HUM. MOL. GENET., vol. 3, 1994, pages 1041
VAN VLIET ET AL., MOL. THER., vol. 14, 2006, pages 809
WEBER ET AL., PROC. NATL. ACAD. SCI. USA, vol. 99, 2002, pages 6222
WENSEL ET AL., VISION RES., vol. 45, 2005, pages 3445
WENSEL TGGROSS AKCHAN FSYKOUDIS KWILSON JH: "Rhodopsin-EGFP knock-ins for imaging quantal gene alterations", VISION RES, vol. 45, 2005, pages 3445 - 3453, XP005192761, DOI: 10.1016/j.visres.2005.07.016
YIN ET AL., IOVS, vol. 52, no. 5, pages 2775
YOKOYAMA ET AL., EXP EYE RES., vol. 55, 1992, pages 225
YOUNG ET AL., OPHTHALMOL. VIS. SCI., vol. 44, 2003, pages 4076
ZHONG LLI BMAH CSGOVINDASAMY LAGBANDJE-MCKENNA M ET AL.: "Next generation of adeno-associated virus 2 vectors: point mutations in tyrosines lead to high-efficiency transduction at lower doses", PROC NATL ACAD SCI U S A, vol. 105, 2008, pages 7827 - 7832, XP002493284, DOI: 10.1073/pnas.0802866105
ZOLOTUKHIN ET AL., GENE THER., vol. 6, 1999, pages 973

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